Biotechnology

The origins of biotechnology date back at least 10 000 years

Describe the origins of biotechnology in early societies who collected seeds of wild plants and domesticated some species of wild animals
Biotechnology began 10 000 years ago when humans changed from nomadic hunter gatherers to farmers Occurred in a few sites at around the same time South East Asia, the Fertile crescent of the Middle East and in south central Mexico o Selected varieties of seeds that produced better quantity and quality of crops and cultivated them o Domesticated wild animals kept certain species close to their homes and allowed them to interbreed

Explain why the collection of seeds and breeding of animals with desired characteristics, could be described as early biotechnology
Humans started to harvest wild seeds and fruits. They found if they set seeds themselves and cared for growing plants, they would be assured of a dependable food source o By collecting seeds from plants with desirable characteristics, they intervened in process of natural selection and humans were now the selecting agents, controlling the genetic makeup of next generation plants Similarly, by choosing animals such as cattle with characteristics they favoured (placid nature), keeping close to homes and allowing them to interbreed provided convenient food source and source of other products (hides, leather and wool for clothing) o Genes of domestic stock different to wild animals and by using living organisms to make products for humans first attempts at biotechnology

Describe the changes in one group of animals and one group of plants as a result of artificial selection of characteristics suitable for agricultural stock
Cattle: Most of our modern species of cattle have arisen by artificial selection o Archaeological and paleontological evidence shows that humans began to keep and breed cattle about 6000BC (8000 years ago) o Believed to have occurred by the domestication of a wild ancestor called the auroch The auroch were large and hairy to enable them to survive harsh climates and evade predators o On domestication, cows and cattle changed in size they became smaller Humans considered characteristics such as high milk yields and good muscle mass to be more important and so chose to interbreed animals with these characteristics o Smooth haired animals became more common (domestication assured them shelter)

Use available evidence to describe the changes in a species of grain or animal as a result of domestication and agricultural processes Process information to outline an ancient Australian Aboriginal use of biotechnology
Biotechnology applied in bush medicine Aborigines had a unique method for treating burns using the outer silk casing of a caterpillar o Caterpillar makes a cone shaped web or nets into which it defecates Aborigines carefully removed the caterpillar from the web and treat the burns with the web o Something in faeces stimulated healing of the burn Domestication of wild dingoes o Used to sniff out certain animals such as the koala when they were sleeping in the day time

Biotechnology has come to be recognised as the use of living organisms to make or modify a product, to improve plants or animals or to utilise microorganisms for specific uses
Outline the key events that led to the use of biotechnological practices, including: yeast in the manufacture of bread yeast and fermentation for alcohol production the use of other micro organisms for the manufacture of yoghurt and cheeses

Yeast in the manufacture of bread: Egyptian tombs show evidence of bread making from around 2000 BC o Since techniques are quite modern, we presume bread making started perhaps 10 000 years ago Wild yeast naturally finds its way into dough, so first leavened bread probably resulted by accident o Yeast ferments sugars to produce carbon dioxide, thus making dough rise Babylonians and Egyptians independently learnt to use old, uncooked leavened dough called starter dough around 1800 BC to speed up fermentation in new batches of dough Analysis of bread samples from around 1500 BC show Egyptians were using a yeast strain called Saccharomyces winlocki to help make baking a more consistent process Today, pure strain of Saccharomyces cerevisiae is almost universally utilised to ferment bread while bacteria used to produce acids that give its unique flavour Yeast and fermentation for alcohol production: Beermaking seems to have begun in Egypt around 6000 BC o Barley was placed in earthenware vessels and buried until it germinated germinated barley is called malt and contains sugars and other compounds that help give beer its flavour o Barley was removed, crushed and made into brewers dough partly baked and afterward soaked in water and at first natural yeasts carried out fermentation however carefully selected yeast were used later o Process modified to introduce heating of malt more efficient way of releasing sugars then yeast added to begin fermentation Winemaking is fairly simple grapes contain natural sugar and natural yeasts grow on their skin o Method was little altered in 5000 years o Several yeasts may be present but only Saccharomyces cerevisiae can survive the higher concentrations of alcohol and produced In ancient times, wine was filtered and stored in earthen ware or animal skin containers; today it is bottled The use of other micro organisms for the manufacture of yoghurt and cheeses: Rock drawings dating back to nearly 10 000 BC show cattle being milked o Cheese would have been made some time after that Improvements in cheese making occurred over the next few thousand years milk was stored in containers and stomach enzymes were deliberately added as it caused milk to sour o Process was sped up through heating o Stomach enzymes cause curd to precipitate and it is separated from remaining watery solution called whey curd removed and dried o During ripening process, moulds often grown on surface giving it a distinct flavour Chinese were using lactic acid producing bacteria (Lactobacillus) in yoghurt making by 4000 BC o Semifluid fermented milk that has a smooth texture but slightly sour flavour because of its lactic acid content

Made in traditional manner by boiling milk in an uncovered pot to evaporate some of the water After cooling, some yoghurt from previous batch is added to boiled milk to introduce bacteria again

Plan, choose equipment or resources, perform a first hand investigation to demonstrate the use of fermentation processes in bread or alcohol production
Method: 20ml of fruit juice was measured with a measuring cylinder and poured into a 250ml conical flask. 1 gram of yeast was added into the solution. The flask was stoppered with a delivery tube. The other end of the tube was inserted into a test tube of limewater The equipment was left in a warm place for two days and changes in the limewater was observed The mixture was left for another 24 hours and then filtered. The filtrate was added into a distillation flask and distilled. The temperature at which a distillate first formed was recorded. The temperature was then monitored to not pass this point. After sufficient distillate collected, its odour was recorded and flammability tested by pouring the liquid into an evaporating basin and lighting it with a matchstick Result: Lime water went cloudy o Calcium carbonate had formed, indicating carbon dioxide was formed from the reaction and therefore confirmed fermentation was taking place There was froth and bubbles which indicated fermentation was also taking place The temperature at which distillate began to form was around 85 degrees Celsius (boiling point of ethanol) The distillate caught on fire, indicating alcohol was produced

Classical biotechnology exploited knowledge of cell biochemistry to produce industrial fermentation procedures
Describe the expansion of fermentation since the early 18 th century to include the production of several organic compounds, including glycerol, lactic acid, citric acid and yeast biomass for bakers yeast

Since the early 18th century major discoveries about the biology and chemistry of fermentation and distillation made it possible to produce cheaper alcohol on a large scale. Over the next 200 years other organic compounds were fermented by using different carbohydrates and micro-organisms. These products included glycerol, lactic acid, citric acid and yeast biomass.

Lactic acid (1880) Milk sugar lactose is fermented to lactic acid by many types of bacteria (commonly Streptococcus thermophiles/ Lactobacillus lactis) o Produce products such as cheese, acidifying foods, dyeing textiles, electroplating and manufacturing plastics Today, lactic acid is made from petroleum Glycerol Glycerol was needed during WW1 to make explosives such as nitroglycerine o At the time, US glycerol was a by product of the manufacture of soap Germany lacked materials to produce glycerol from soap production and therefore had to make it by fermentation o When alcohol is produced by normal fermentation, yeasts also produce small amount of glycerol. o Addition of sodium hydrogen sulphite or sodium bisulfite interferes with the biochemical pathway between glucose and ethanol and forces production of glycerol: up to 20 30% of sugar used can be converted into glycerol Many uses of glycerol: solvent, a sweetener, antifreeze mixtures, in medicine and in the production of dynamite

Citric acid There are several moulds that can convert sugars into citric acid. Most commonly, fungus Aspergillus niger is used on submerged fermentation of glucose o Surface production 1923 o Submerged production 1930s Submerged production sterile medium and mould are added to a large tank where it is constantly mixed and a supply of sterile air bubbled through Used in food industries as a flavour enhancer Bakers yeast Stock strain is added to a food medium mainly molasses and pH is kept around 4 5, helping to retard bacterial growth Yeast require aerobic conditions to grow and medium is aerated while yeast grows Yeast is harvested by washing cells and then separating it by using centrifuges. A filter press is then used to squeeze out the remaining water before the yeast is packed

Describe strain isolation methods developed in the 1940s

During strain isolation, a population of cells is grown from one or more cells that have a desired characteristic pure cultures of the particular micro organism are then obtained o Best strains are often found by accident penicillin Planned screening methods have also been used to find new sources of useful materials o Soil, hot springs, salt lakes, wood bark and roots useful microbes are often found o Screening is carried out to test microbes effectiveness against disease causing bacteria, viruses and tumours Example: Microorganisms are streaked across an agar surface with a loop and as it is streaked, more and more bacteria are rubbed off until individual separated organisms are deposited on the agar o After incubation, area at beginning of streak pattern will show confluent growth while area near end of pattern should sow discrete colonies This microorganism is screened and bacterial colonies that have desired characteristics are isolated and used to inoculate another petri dish o This process is continued until a pure culture of bacteria is established

Describe, using a specific example, the benefits of strain isolation methods used in biotechnology in the 20th century
Strain isolation methods, in which pure cultures of a particular micro organism are obtained, have been of great use to mankind over the last century Most notable instance is the discovery and isolation of fungus Penicillium by Alexander Fleming in 1929. Fleming accidentally found that staphylococcal colonies he was culturing did not grow in the vicinity of Penicillium colonies that had contaminated his agar plates o Concluded that fungus was producing substance that destroyed cell walls of bacteria and named substance, Penicillin o Using rudimentary strain isolation techniques, produced sub cultures of Penicillin to isolate this antibiotic More sophisticated isolation techniques was used by Howard Florey who produced a process for large scale production of penicillin was developed by Howard Florey use for casualties in WW2 Modern strain isolation procedures have proved invaluable in epidemiological work o Allow pathologists to diagnose specific infectious diseases in humans and also to locate original source of these diseases o E.g. food poisoning disease botulism can be confirmed by isolating pure cultures of bacterium Clostridium botulinum from food samples and stools of afflicted people

Identify that developments in the 1950s led to biotransformation technologies that could produce required organic compounds such as cortisone and sex hormones
Biotransformation involves use of micro organisms or enzymes derived from living organisms to create specific chemicals from precursor molecules

o Refers to the modification of a chemical structure by a living organism Examples of biotransformation technologies include the production of hormones such as cortisone, hydrocortisone, prednisolone and testosterone using microbes ore enzymes

Gather and process information from secondary sources to: identify and describe a named industrial fermentation process identify the micro organism used in the fermentation and the products of the fermentation outline the use of the product of the fermentation process use available evidence to assess the impact of the use of the fermentation product on society at the time of its introduction

Identify and describe a named industrial fermentation process Conditions of normal fermentation of sugar are changed to produce glycerol sodium hydrogen sulfite or sodium bisulfite is added which interfere with the biochemical pathway between ethanol and glucose and forces the production of more glycerol (up to 20 30% of the sugar used can be converted into glycerol, with fermentation taking 2 3 days) Undertaken in bio reactors (fermentation tanks) which can hold several thousand litres of the fermentation mixture o Contains nutrients, stirrers, pH and temperature controls and aeration devices Identify the micro organism used in the fermentation and the products of the fermentation Yeast used Saccaromyces cerevisiae Sodium hydrogen sulfite or sodium bisulfite added during fermentation process Outline the use of the product of the fermentation process To produce explosives (dynamites) Solvent, sweetener, antifreeze mixtures, in medicine Use available evidence to assess the impact of the use of the fermentation product on society at the time of its introduction At the time U.S. glycerol was a by- product of the manufacture of soap. In contrast, Germany lacked the materials to make glycerol from soap production and therefore had to make it by fermentation o Although this process is no longer used, it serves to show how the modification of fermentation conditions can alter the product o Extremely significant impact because glycerol was then used during WW1 to make explosives (such as nitroglycerine), resulting increasing the duration of the war and resulting the loss of many lives as a result of the development

Process and analyse information from secondary sources to demonstrate how changes in technology and scientific knowledge have modified traditional uses of biotechnology, such as fermentation
Changes in technology/scientific knowledge Discovery and understanding of yeasts Impact on traditional uses of biotechnology Change from the sourdough breads that rely on wild yeasts to the deliberate incorporation of specific cultures of yeast in bread making Bacteria that caused harmful fermentation could be eliminated Safer, more consistent products, large scale production possible Widespread production of products such as lager that was once restricted to the icy lake areas in parts of Europe Better quality, more consistent wines

Pasteurs discovery that bacteria could be killed by heating Knowledge and ability to isolate and grow pure strains of micro organisms Development of refrigeration

New technologies for crushing and filtering wines

Cell chemistry is utilised in biotechnology

Outline, simply, the steps in the synthesis of a protein in the cell including: The difference between DNA and RNA The production of messenger RNA The role of transfer RNA The formation of the polypeptide chain(s) The formation of the protein from polypeptide chains
DNA Nearly always double stranded Thymine, Adenine, Cytosine and Guanine Ribose Nucleus only RNA Usually a single stranded molecule Uracil, Adenine, Cytosine and Guanine Deoxyribose Nucleus + cytoplasm

Difference between DNA and RNA Shape Nitrogen bases Sugars Location

Production of messenger RNA Enzyme, RNA polymerase, binds to a part of the DNA called the promoter and the DNA unzips that is the DNA unspirals, hydrogen bonds between the two strands break, and the strands separate over a short length, just in that part of the DNA that holds the gene to be used Transcription of the genes occurs, controlled by the enzyme RNA polymerase. The single strand of DNA acts as a template and RNA nucleotides are assembled, forming a complementary single stranded mRNA molecule o Sequence of nucleotide bases on mRNA molecule is complementary to the DNA coding strand, except that it has U instead of T Role of transfer RNA Particular set of RNA molecules called transfer RNA (tRNA) is involved in translation tRNA is synthesised in the nucleus and pass through membrane at same time that mRNA is being produced There is a triplet of tRNA bases that codes for a specific amino acid mRNA goes to the ribosomes on the endoplasmic reticulum and the triplets of transfer RNA picks up their corresponding amino acids in the cytoplasm and brings them to the mRNA. Translation occurs where ribosomes move along the mRNA molecule and attach tRNA molecules by pairing bases of tRNA anticodons with complementary triplets of bases on the mRNA (codons) Formation of polypeptide chain(s) Enzymes join the amino acids attached to the tRNA together to form a polypeptide chain o Continues until polypeptide chain is finished and ribosome has rolled along almost the whole of the mRNA chain AT the end, mRNA will have a codon for stop and this will cause enzymes that terminate the polypeptide chain to cleave it (break it) from the ribosome. Formation of the protein from polypeptide chains Some proteins consist of one single polypeptide chains, others consist of more than one Polypeptide chain may be joined by one or more other polypeptides; they are further processed and folded into their correct shape, forming a protein o Different chemical attractions and repulsions between the side chains on the 20 amino acids along the polypeptide chain can cause it to twist and fold in many different ways

Plan and perform a first-hand investigation to test the conditions that influence the rate of enzyme activity

Modern biotechnology includes recombinant DNA technology

Describe the three essentials of gene manipulation as: cutting and joining DNA monitoring the cutting and joining transforming hosts, such as bacteria, with the recombinant DNA

Cutting and joining DNA DNA that is required is cut from a long strand of DNA (a chromosome), using restriction enzymes A circular piece of DNA, called a plasmid, that is present in bacterial cells, is removed from the cells and is cut open using the same restriction enzyme The gene is then mixed with the bacterial plasmids in a test tube. Because they have been cut with the same enzymes, the cut ends of the plasmid and the end of the gene match o Sticky ends match Enzyme: DNA ligase is used to stick the ends together Monitoring the cutting and joining When bacteria are cultured so that cells multiply, no one can see whether there has been exactly one recombined plasmid in each new cell; or whether some cells have lots of the new plasmids and some cells have none. It is also possible some small, inactive parts of the host DNA have been inserted into the plasmids o Scientists need to monitor which bacteria contain the new, complete plasmid Usually marker genes are used to help monitor progress of research with gene of interest commonly a gene for antibiotic resistance is used DNA from marker gene for antibiotic resistance is inserted into plasmid along with gene to be cloned becomes part of the new, recombined DNA When modified bacteria have been cultured, antibiotic resistant gene will also be cloned alongside it Sample of the cultured bacteria is then streaked onto an agar plate (or added to a liquid medium in a tube) that contains that particular antibiotic o Only bacteria containing new gene will grow, as they also carry gene that makes them resistant to that antibiotic o Scientists left with cultures they require Researchers usually use two different marker genes and choose colonies that survie both processes IN other cases, gene probe with a fluorescent or dye molecule is added Transforming hosts, such as bacteria, with the recombinant DNA Technique for inserting recombinant plasmid back into the bacterium is called transformation Organism receiving the newly modified genome is called the host This can be done by combing the recombinant DNA with a host (likely bacteria) in a test tube with calcium chloride high conc. of calcium ions make membrane of bacteria more porous o DNA molecules in solution have an overall negative charge due to the phosphate molecular groups in their backbone. The phospholipids of membranes of all cells also have negative charges there are pores in the membrane that open or close to allow essential molecules through o By mixing the plasmids and bacteria in solution with high conc. of calcium chloride at 0 degrees, the positive calcium ions slow their movement in solution and bind to some of the proteins and to the bacterial membrane, giving the pore an overall positive charge and attracting negative DNA that is in the plasmid If solution is heated quickly enough to about 35 degrees, the pores open upand plasmid enters bacterial cell Some chloride ions enter host cell also, causing surrounding water to enter by osmosis so cell swells, helping pores to open up

Describe the following recombinant DNA techniques used in biotechnology, including: gene splicing using restriction enzymes and ligases to produce recombinant DNA polymerase chain reaction to amplify or modify DNA sequences use of DNA vectors and microinjection for carrying genes into nuclear DNA in the production of transgenic multicellular organisms

Gene splicing using restriction enzymes and ligases to produce recombinant DNA: Gene is cut out of the chromosome of one species using a restriction enzyme Circular piece of DNA known as a plasmid, present in bacterial cells, is removed from the bacterial cells and cut open with the same enzyme Cut out gene is mixed with bacterial plasmids in test tube o Because they have been cut with same enzymes, cut ends of plasmid and ends of gene match. Therefore a plasmid recombines with the gene spliced into it Sticking together of the plasmid with the gene inserted into it is catalysed by the enzyme DNA ligase Polymerase chain reaction to amplify or modify DNA sequences DNA polymerase is a reaction in which DNA polymerase is used to produce more DNA from very small amounts of DNA or from impure samples of DNA o DNA to be amplified is mixed with deoxyribonucleotides, thermal stable DNA polymerase called Taq polymerase and DNA primers Three steps are involved in PCR o Denaturation o Annealing o Extension Denaturation DNA is heated to 95 98 degrees Celsius to separate the two complementary strands into single strands o Denaturation does not damage DNA as it does proteins (enzymes) Annealing mixture is cooled sufficiently to allow the DNA primers to anneal or stick onto single strands of DNA at locations where they are complementary Extension Taq polymerase then synthesises the complementary strand of DNA, using the primer as the starting point Strands are then reheated and process is repeated so fast that in five minutes the required DNA sequence has been copied. By about 20 25 cycles, there is a massive amplification of the DNA sample for further testing and analysis Usefulness: Criminal cases in which very little DNA is available to identify the criminal/s. Use of DNA vectors and microinjection for carrying genes into nuclear DNA in the production of transgenic multicellular organisms DNA vectors: DNA vector commonly used to transfer genes into plants is a plasmid (Ti plasmid) o new gene/s (from another species) is inserted into the plasmid using recombinant DNA technology. Either plasmid is transformed into the bacteria, which are then introduced into the plant cells or directly inserted into the plant cell Free plasmids can easily invade plant cells and plasmid DNA will merge with the plant chromosomes When plant grows, it will express the new characteristic and will pass it onto its offspring Another common DNA vector are viruses o Viral vector modified so that it will not replicate or cause disease in the target cells of host embryo o Gene of interest is incorporated into the viral genome (using recombinant DNA technology) and virus is then used to infect an early stage embryo or pronuclear embryo

Viral vector binds uniformly to the embryonic cells and acts as a vehicle to allow transfer and integration of the transgene into the host cell

Microinjection: Involves placing of new DNA into an animal genome by injecting it into the nucleus of a fertilised egg (zygote) Very fine glass needle is used to inject pure sample of DNA from another species (or sometimes from another individual of same species) into cell o Microscope can be used to monitor the process Microinjected eggs are then transferred into the uterus of each of specially chosen surrogate mothers

Process and analyse secondary information to identify that complementary DNA is produced by reverse transcribing RNA or the polymerase chain reaction
Another method for amplifying DNA is by reverse transcription, that is, transcription from RNA to DNA Method: 1. Double stranded DNA containing introns and exons 2. Transcription of the DNA occurs in the cell to form mRNA 3. Introns are removed from the mRNA by restriction enzymes and it is extracted from the cell 4. Enzyme reverse transcriptase is used to produce a single strand of complementary DNA from the mRNA 5. In the presence of DNA polymerase, a double strand of the complementary DNA is then formed Examples: Retro viruses such as AIDS virus have only RNA and a reverse transcriptase enzyme o Enter host cell and cause it to manufacture viral DNA by reverse transcription Employed in laboratory to produce DNA sample for which a corresponding RNA sample is present using a reverse transcriptase enzyme

Perform a first hand investigation to extract and identify DNA from a suitable source Process information to produce a flow chart on the sequence of events that result in the formation of recombinant DNA Gather and analyse information to outline the purpose of a current application of transgenic technology, naming the organism and gene transfer technique involved
Purpose of Bt cotton plants In 1990s, CSIRO scientists in collaboration with US company Monsanto extensively trialled use of Bt cotton transgenic species Caterpillar of the Helicoverpta zea moth destroys hundereds of millions of dollars worth of cotton each year Bt cotton plants were genetically modified and contain the Bt gene which code for production of toxic protein in an inactive form that is harmless to humans/ most animals and insects, however, when protein eaten by caterpillar - converted by the digestive system into active form that kills the insect o Gene called Bt originally taken from soil bacterium Bacillus thuringiensis Bt cotton plants are used as it reduces the need to use pesticides to kill caterpillars which is better for the environment and reduces development of pesticide resistance in caterpillars Technique involved Scientists cut normal cotton seedlings, place them on solid growth medium grow into calluses. After about six weeks, callus cells are transferred to liquid medium, given hormones to induce them to grow into cotton plant embryos Bt gene is extracted from bacterium Bacillus thuringiensis using restriction enzymes Bt gene is transferred to cotton plant embryos o Using a vector bacterium Agrobacterium tumefaciens that is able to inject genes into other cells Cotton plant embryos are dipped in solution of mixture vector, and extracted Bt genes o Vector bacteria inject Bt genes into cotton cells Embryos containing Bt genes are grown in tissue culture and placed on another solid medium and germinated into small plants transgenic species

There are many applications and areas of research in biotechnology

Outline one way that forensic scientists can use DNA analysis to help solve cases
DNA fingerprinting is one of a number of uses of DNA technology o Used by forensic scientists to help solve cases DNA fingerprinting detects with some accuracy the blood, tissue or semen DNA of an individual and identifies the individual in either criminal or paternity cases Process of comparing DNA involves using restriction fragment length polymorphisms Restriction enzymes are used to cut DNA into fragments at precise points Pieces of DNA are run through an electrophoresis gel to separate the lengths of fragments The fragments are collected onto a screen or sheet of special material in a blotting process One or more of the fragments is visualised with a probe. The probe is really a molecule of single stranded DNA that is complementary to the sequence of one of the fragments of DNA and is either radioactive or fluorescent The probes bind with the appropriate fragment and thus identify it Pattern that is obtained can then be compared to patterns obtained from DNA obtained at the crime scene Uses of DNA fingerprinting Method can be used because DNA base sequence varies with individuals For most analyses, DNA regions that are known to be highly variable from one individual to another are chosen o Not all DNA fragments are compared result will indicate a level of probability that suspect may actually have been at site and left the tissue/blood/semen o The more regions analysed, the higher indication of probability if individual was present or not DNA fingerprinting alone does not prove a person guilty but can be used to prove a person innocent

Identify data sources, gather, analyse and process information to present one case study on the application of biotechnology in each of the following: medicine animal biotechnology aquaculture

these case studies should: give details of the process used identify the organism or tissue involved describe the outcome of the biotechnological process evaluate the efficiency of the process and discuss advantages and disadvantages associated with either the product or the process

Application of biotechnology in medicine production of synthetic insulin Process used: Gene for making insulin is cut out from nucleus of a human pancreas cell using a restriction enzyme Plasmid is removed from bacterial cells and cut open with same enzyme The insulin gene is mixed with the bacterial plasmids and because they have been cut with the same enzyme, the cut ends of the plasmid and ends of genes match. The sticking together of plasmid with gene inserted into it is catalysed by enzyme DNA ligase The plasmid, containing recombinant DNA is re inserted into Eschericia coli (E.coli) bacterial cells. Once introduced, bacteria are grown in large tanks in manufacturing plants at optimal temperatures o Millions of bacteria replicate by binary fission and express the genes

Once desired amount is produced, cell wall is broken open to extract the DNA. The DNA mixture is the purified, resulting in insulin chains remaining

Identification of the organism or tissue involved: The organism (bacteria) in which plasmids are removed from and re- introduced into are known as Eschericia coli Description of the outcome of the biotechnological process: Outcome is synthetic insulin which can be used to treat people with diabetes mellitus (Type 1 diabetes), a disease where body cannot maintain normal blood glucose levels due to insufficient insulin production by the pancreas o Symptoms are eventual kidney failure, damage to vision and restrictions of blood flow to limbs Advantages and disadvantages associated with either product or process: More than 50 years ago, insulin was extracted from pancreas of calves and pigs o Due to small differences in calf and pig insulin from human insulin, some allergic reactions were observed One great advantage of synthetic insulin is that it is 100% insulin and hence less antigenic o Fewer abnormal reactions with hormone as there is no triggering of immune system Process of producing insulin is advantageous as the product can be produced very quickly in vast amounts as bacteria has a fast reproduction rate with fewer resources needed than complex animals. It is less costly than previous means of producing insulin as space needed to support specialised bacteria colonies is significantly smaller than needed to raise livestock Disadvantage is insulin has to be constantly injected by patient almost every day to treat the diabetes Disadvantage of synthetic insulin produced is initially there was an increase of more than 3 times the hypoglycaemic complications compared with animal insulin o Hypoglycaemia characterised by abnormally low levels of sugar in blood due to too high insulin intake resultant symptoms of weakness, shakiness, nervousness and fainting Complications have decreased but remains disadvantage as product is faced with continuous doubt and scrutiny

Evaluation of efficiency of process: This process in producing synthetic hormone, insulin, is very efficient due to the exponential rate of reproduction of E. Coli bacteria. The bacteria reproduces by binary fission, where one bacteria splits into two, the two splits into four and the four splits into eight and so on. Therefore, as the bacteria reproduces exponentially and expresses the genes, insulin is produced very quickly and in large amounts. The E. Coli bacterium also reproduces asexually, allowing it to be managed much more easily, making the process cost effective. This process is also very efficient in meeting the increasing demands of synthetic insulin. Although there are some disadvantages of the product itself, such as some reported cases of hypoglycaemia complications and the need to inject the product into the body, overall these can be negated by the ability of synthetic hormone to improve the diabetics quality of life (by reducing risk factors such as constrictions of blood flow and kidney failure). The level of people with diabetes mellitus is increasing at an alarming rate, however the process is efficient in matching these demands by being able to generate a large yield of synthetic insulin in a short amount of time. The overall process of the production of synthetic insulin is therefore very efficient. Application of animal biotechnology Details of process used: The immune system of a mouse or rabbit is triggered by injecting a specific antigen for which the antibody is required The lymphocytes (B cells) produce an antibody to attack the specific antigen. These B cells are isolated from the spleen of the animal

The lymphocytes are fused with rapidly growing special tumour cells to form cells called hybridomas. The hybridomas are cultured, producing monoclonal antibodies in great quantities o B cell part of the hybridoma produces the antbiodies and the tumour cell part divides rapidly to produce more cells, which produce more antibody. Hence high yield Monoclonal antibodies produce is extracted and purified used to detect or treat diseases caused by the antigen that originated them

Identification of the organism or tissue involved Organism involved in producing B lymphocytes can be either a mouse (Mus musculus) or rabbit Tumour cells that are fused with isolated lymphocytes are normally myelomas Description of the outcome of the biotechnological process Monoclonal antibodies can be used to detect pregnancy. Females produce a protein hormone, human chorionic gonadotrophin (HCG). MAb, specific for this hormone can be produced by the process and added to a urine sample of a female. If HCG is present, antibody binds with it and enzymes associated with HCG produce a colour with a reagent, which identifies pregnancy MAbs used to diagnose infection with HIV, cancer treatment, used to neutralise snake, spider venoms and bacterial toxins Advantages and disadvantages associated with either the product or the process Before production of MAbs by this process, it was produced by isolating B cells from immunised animal and then culturing it. This process is very advantageous due to inclusion of myeloma. By fusing B lymphocytes with tumour cells for rapid division, the rate of production and yield of monoclonal antibodies has increased dramatically o Monoclonal antibodies can be produced in large amounts within a short period of time Advantage is the stability of monoclonal antibodies within the human body. Recent advances in antibody technology has allowed monoclonal antibodies to remain stable in body for long periods of time making it an effective cancer treatment o Able to remain in blood for long periods of time, circulating and targets cancer cells for destruction by immune system Disadvantage ability of monoclonal antibodies to correctly destroy antigen for which they were produced. MAbs are able to precisely identify and locate their intended targets, however some issues arise in their ability to neutralise or destroy the antigen o They either attack human cells, damaging healthy tissue or do not act when they arrive at intended target Disadvantage some MAbs produced when introduced into human body can be recognised as foreign proteins. Body produced antibodies that attack the MAbs and eventually neutralise them o In some cases, MAbs become ineffective in targeting a specific antigen as body acts to destroy them

Evaluation of the efficiency of the process The process in producing monoclonal antibodies is highly efficient due to the myeloma cells trait of dividing endlessly while the B cell produces the specific type of antibody. The constant dividing of the tumour cell allows the production of the antibody to be exponential in nature and also produces a high yield of monoclonal antibodies compared to the old process in culturing the isolated B - lymphocytes. There are a variety of uses for monoclonal antibodies including pregnancy testing, cancer treatment, assessing the extent of damage to heart muscle, neutralising venoms and much more. Due to the variety of uses of monoclonal antibodies, the demand for this product is high and the process is very efficient in meeting this demand due to the high abundance at which monoclonal antibodies can be produced. However there are some problems, which act to reduce the efficiency of the process. During the process, there are some cases of hybridoma culture contamination and the process is expensive and time consuming to set up. Yet

these disadvantages are quite minimal and are outweighed by the advantages and therefore the process in the production of monoclonal antibodies is deemed very efficient. Application of biotechnology in aquaculture Details of process used: - selective breeding: A yabby population is established in a marine environment such as a pond or dam area with some in built infrastructure Selective breeding process is utilised where yabbies that possess favourable characteristics such as faster reproduction rates and larger tails (containing large amount of edible meat) are selected and grown, so that animals with these characteristics become more prevalent within population of marine environment o Process involves discarding individuals in selection process so that only favourable characteristics are passed onto successive generations During process, yield can be increased by supplementing yabbies with higher quality feeds, increasing temperature or increasing marine environment Identification of organism or tissue involved: Organism involved in process (farmed) is the Australian freshwater crayfish, the yabby (Cherax destructor) Description of the outcome of the biotechnological process Yabbies become larger and contains more edible meat than those not selectively bred o Sold to pet stores and distributors of restaurants (popularity for yabbies to be included in gourmet restaurant menus have soared recently) Yabbies can be harvested continuously by leaving a portion within the marine environment in successive harvests in order for yabbies to reproduce Large yield of yabbies is produced, increasing amount of yabbies able to be sold to the market and hence increasing the profitability of process Advantages and disadvantages associated with either the product or process: Advantage of process offers sustainability. After yabbies are harvested, portion is left within marine environment so that they can reproduce for the next harvest o Environmentally sound as yabbies do not have to be continually collected from their habitats Advantage of process it improves population of yabbies in its characteristics over a period of time. Through selective breeding, favourable characteristics become more prevalent within population. Therefore they can be sold at higher price, increasing profitability of the process Disadvantage of process yabbies in the farm have a high mortality rate (up to 95% in some cases) because problems of overcrowding, high stress levels and limited feeding in environment results in their deaths Even for yabbies that survive, many have broken claws/legs unsuitable to be sold to market o Therefore process considered inefficient to an extent Disadvantage of process method of farming requires a great deal of habitat development where five areas of environmental conditions need to be addressed (air, food, temp, water quality and shelter) To meet criteria with addition of pond construction and dam maintenance, this type of marine farming can be quite costly to set up initially

Evaluation of the efficiency of the process: On one hand, the process could be deemed inefficient due to the high mortality rate of the yabbies, leading to a low yield and due to the high cost in setting up the marine environment for the yabbies to live and reproduce. However these disadvantages of the process can be negated by feeding the yabbies with high quality food, increasing the area where the yabby population is grown and by taking out the yabbies when they are young to be grown individually in bottles to decrease overcrowding and stress. The cost in setting up the environment is also normally negated in the long term when yabbies grown from the marine environment are continually sold to the market.

The process also contains various advantages such as sustainability whereby yabbies can be continuously grown without interfering yabby populations in other habitats. The process also allows the presence of yabbies with desirable characteristics to increase, allowing for higher quality yabbies to be sold and in turn, increasing profitability of the process. Therefore the process is quite efficient not only due to the advantages described but because of the ability to negate the disadvantages identified through various techniques as discussed above.

Ethical issues relevant to the use of biotechnology are important and need to be considered
Explain why different groups in society may have different views about the use of DNA technology
A scientist, a lawyer, a biotechnology company, a patient, a farmer, a consumer each may have a different opinion about the development and use of DNA technology

Scientists: Views on DNA technology are generally positive because they are usually strictly concerned with the practical development of the technology o Mindset that anything new that can be done should be done Lawyers: Grappling with many new advances in gene technology They earn quite large amounts of money from the development and implementation of laws associated with gene technology Biotechnology companies: Very positive about DNA technology reason why many of these new industries exist Employees depend on DNA technology for jobs Boards of directors strive to have the biotechnology company listed on the stock market, so providing more funds for research and new products Profit motive is always there o Some companies choose to keep trade secrets about how a product or products of DNA technology are prepared Patient: For many patients, DNA technology shows great promise A patient who is suffering from a disease that can be treated or cured by DNA technology is usually desperate to avail herself or himself of the technology Consumers: Mixed feelings about DNA technology o It all seems too fast and complicated Some consumers are wary about the hazardous properties of genetically modified organisms and in genetically modified foods Farmers: Mixed feelings about DNA technology On one hand, recombinant vaccines such as TickGARD improve survival and growth of cattle, increase yield and avoid use of chemicals in the environment On the other hand, many genetically modified crops are still the great unknown and may pose a threat to natural crops Agribusinesses also take the bulk of the profit o Some farmers see this as taking away their autonomy

Identify and evaluate ethical issues related to one of the following:

Development of genetically modified organisms (GMOs) Animal cloning Gene cloning

In Australia, as in other countries, there is a great debate about GMOs, especially GM foods

Ethical issues: Transfer of genes from animals whose flesh is forbidden into other animals may be offensive to certain religions o Jews and Muslims do not eat pork and in some cases pig genes are transferred into sheep Transfer of animal genes into plants may be of concern to vegetarians, especially vegan Recombinant DNA may be taken up by non target, organisms. These unintended GMOs may have the potential to become pests or cause disease Questions of concern arises: o Could consumption of GM food cause disease due to a toxin or an allergen present in food due to some contaminating genetic material from the donor organism or a vector? o Could the other characteristics of t he GMO be changed by introduction of the new gene/s? Benefits: Providing easily grown, high yield organisms for food to relieve hunger Use of GM foods will result in the use of less chemicals such as herbicides and pesticides Not just the quantity of the food but also better quality foods are produced Vaccines can be produced in plants for better human health outcomes Produce may reach consumer in better condition with less spoilage

Evaluation: Legislation to control the production and release of GMOs varies in different countries o Controls are usually rigorous and strictly enforced The benefits of GMOs seem to outweigh the ethical issues related to it. Some may argue that it is unethical to develop GMOs if it has the potential to do good

Use available evidence to identify and discuss ethical and social issues associated with the use of biotechnology