Technology - Tortora - Rio Salado - AZ Question Answer How can microorganisms & plants be used as "factories" to By inserting genes into cells by recombinant DNA produce unnatural tech (rDNA) - genetic engineering chemicals/products? Another name for recombinant genetic engineering DNA (rDNA) technology. A plasmid or virus used in genetic engineering to vector insert genes into a cell. DNA vectors are often called __. gene-cloning vectors, or cloning vectors. Techniques used to modify a gene in a specific Site-directed mutagenesis. location to produce the desired polypeptide. Site-directed mutagenesis can be make a specific change in a gene. used to __. Special class of DNA - cutting enzymes that exist restriction enzymes in many bacteria. Restriction enzymes protect a hydrolyzing phage DNA - protected because cell bacteria from __. methylates (add methyl groups). Using restriction enzymes to add a methyl group to methylates cytosines in DNA. What role do restriction enzymes Recognizes & digests (cuts) only are particular play? sequence of nucleotide bases in DNA. They "stick" to complementary stretches of single- What is useful about sticky ends stranded DNA by base-pairing. Same enzymes left by staggered cuts? used on 2 different sources of DNA will splice (recombine) in vitro. (1) restriction enzyme cuts 2 DNA at its particular Order of formation of rDNA site, (2) sticky ends formed, (3) sticky ends bond molecule. w/hydrogen bonds, (4) DNA ligase covalently bonds backbones of DNA pieces to produce rDNA molecules. Because they produce staggered cuts that create Why are restriction enzymes used "sticky ends" that can bond w/other DNA strands to make recombinant DNA? cut by same enzymes. Most important property of a self-replication vector. What does an ori (origin of replication) allow a plasmid to be self-replicating do? vehicles for the replication of desired DNA Vectors serve as __. sequences. Which size of vector is easier to Smaller vectors, because larger DNA molecules manipulate? are more fragile. Most important property of a self-replication vector. What does an ori (origin of replication) allow a plasmid to be self replicating do? vehicles for the replication of desired DNA Vectors serve as __. sequences. Which size of vector is easier to Smaller vectors, because larger DNA molecules manipulate? are more fragile. They are plasmids that are capable of existing in What is special about shuttle several different species & can be used to move vectors? cloned DNA sequences among speces. Vector that can usually accept much larger pieces Viral DNA of foreign DNA than plasmids can. Name some viruses that are used Retro viruses, adenoviruses, & herpes viruses. as viral vectors. Technique by which small samples of DNA can be Polymerase chain reaction (PCR) quickly amplified (increased quantities). PCR can amplify __. small samples of DNA Short pieces of nucleic acid added to help start primers reactions - complementary to ends of target DNA & hybridize fragments. After each cycle of synthesis, DNA is __ to convert all the new heated DNA into a single strand. What enzyme copies DNA in DNA polymerase PCR? DNA polymerase for PCR comes Thermus aquaticus - a thermophilic bacterium - from __. allows for heating by a thermalcycler. PCR __ be used to amplify an cannot entire genome. In genetic engineering, a plasmid transformation is inserted into a cell by __. Procedure during which cells can take up DNA from environment - cells soaked in Ca chloride & transformation made competent - heat added & then they take up DNA. Using electric current to create micro pores in electroportation membrane - cells then absorb DNA through pores. Cell walls removed enzymatically to make bacteria protoplasts vulnerable to experimentation. Protoplasts in a solution will fuse & create hybrid Protoplast fusion cell - can merge species like plant & alge. Microinjection inserting DNA into a cell w/glass micropipette Foreign DNA will only survive if is present on self-replicating vector or is it __. incorporated into a chromosome by recombination. collection of clones containing different DNA gene library fragments. Genes are cloned into vectors restriction enzymes using __. The enzyme reverse transcriptase complementary DNA (cDNA) is used to synthesize __. cDNA is synthesized from __ an mRNA template. A DNA copy of mRNA is reverse transcriptase produced by __. cDNA DNA made in vitro from an mRNA template. The __ method is most common method of obtaining eukaryote cDNA genes. Synthetic DNA is created mechanically by How does cDNA differ from assembling nucleotides as if letters of a keyboard, synthetic DNA. while cDNA contains exons from a mRNA template. If a bacterium received the original plasmid Blue-white screening for clones containing a piggy-backed gene, it will hydrolyze x-gal & produce a blue-colored compound. ID of a colony containing a desired gene by using Colony hybridization DNA probe complementary to that gene. Short, labeled, single strand of DNA or RNA used DNA probe to locate its complementary strand in a quantity of DNA. DNA probes are labeled with? A radioactive element of fluorescent dye. In blue-white screening, white Foreign DNA contains? In blue-white screening, blue no foreign DNA contains? Why is E. coli used in genetic They are grown easily & their genetics are known. modifications. What is one disadvantage of using They are gram-negative & produce endotoxins. E. coli for genetic engineering. Why use yeast cells in genetic They are eukaryotic & may carry plasmids. engineering? gene silencing Defense against viruses & transposons. Target a particular gene - like a virus gene - and RNA interference (RNAi) binds to mRNA & destroys it - silencing the expression of the gene. One technique for genome random shotgun sequencing. sequencing is __. Technique for determining the nucleotide Random shotgun sequencing sequence in an organism's genome. The science of understanding the function of genes bioinformatics through computer-assisted analysis. Science of determining all of the proteins proteomics expressed in a cell. 2 uses of Southern blotting Genetic screening & DNA fingerprinting.