EREDITATE CONSERVAREA CARACTERELOR PARENTALE - SUB INFLUENTA FACTORILOR DE MEDIU

SUPORTUL MOLECULAR AL EREDITATII

MENDEL- 1865MENDEL- 1865EREDITAR EREDITAR notiunea de FACTOR

GRIFFITHGRIFFITH- 1928 efectul transformant al capsulei de diplococus pneumoniae

polizaharidice

AVERY, CARTY, MacLEOD, 1944 ADN FACTOR TRANSFORMANT MacLEOD, MODEL EXPERIMENTAL PE SOARECI INOCULATI CU TULPINI DE DIPLOCOCUS PNEUMONIAE: (VIRULENTE- S, NEVIRULENTE- R) (VIRULENTENEVIRULENTE1) AMESTEC TULPINI R CU PROTEINE DIN TULPINI S
2) CULTIVAREA FORMEI R CU ARN DIN TULPINI S 3) CULTIVAREA TULPINII R CU CAPSULA MUCOPOLIZAHARIDICA DIN TULPINA S 4) CULTIVAREA TULPINII R CU ADN EXTRAS SI PURIFICAT DIN FORMELE S AU APARUT COLONII VIRULENTE CARE INJECTATE LA SOARECI AU PROVOCAT MOARTEA ACESTORA 5) CULTIVAREA FORMELOR VII R CU ADN S DISTRUS IN PREALABIL CU ENZIME- DN-AZAENZIME- DN-AZA- SE OBTIN FORME R NECAPSULATE SI NEVIRULENTE

CONCLUZIE: ADN ESTE SUPORTUL EREDITATII

CARACTERE ADN - IMPLICATIA IN EREDITATE

ARE STRUCTURA SPECIFICA SPECIFICITATE DE SPECIE PRIN ORDONAREA BAZELOR AZOTATE. AZOTATE. ARE CAPACITATE DE SINTEZA ( AUTOREPLICARE). AUTOREPLICARE).

INFORMATIA ADN POATE FI DECODIFICATA SI TRANSMISA ARN SINTEZA DE PROTEINE CARACTERE. CARACTERE. ESTE SURSA DE VARIABILITATE MUTATIE. MUTATIE. PRIN RECOMBINARE SI

ARE O DISPUNERE LINIARA, INFORMATIA DETINUTA FIIND ACCESIBILA. ACCESIBILA.

STRUCTURA ADN LOCALIZAREA CELULARA A ADN : -NUCLEU 98% CITOPLASMACITOPLASMA- MITOCONDRIE 2%

STRUCTURA PRIMARA : MACROMOLECULA CU

GRAD INALT DE POLIMERIZARE.

UNITATEA STRUCTURALA = NUCLEOTIDUL ( BAZA AZOTATA + PENTOZA+ REST DE FOSFAT ANORGANIC) PENTOZA+

DNA Building Blocks

Nitrogenous Base Pentose Sugar Triphosphate

5 Phosphate 3 Hydroxyl

Nitrogenous Base Structure

Pyrimidines Single Ring Bases (T and C) Purines Double Ring Bases (A and G)

NUCLEOZIDUL = BAZELE AZOTATE LEGATE la C1 AL DEZOXIRIBOZEI

Ex: ADENOZINA, GUANOZINA, CITIDINA, TIMIDINA

NUCLEOZIDUL SE LEAGA PRIN C5 AL DEZOXIRIBOZEI DE ACIDUL FOSFORIC POLIMERIZAREA SE FACE PRIN LEGATURI 3- 5 FOSFODIESTERICE INTRE C3 AL DEZOXIRIBOZEI SI POZITIA C5 A NUCLEOTIDULUI URMATOR-

SE REALIZEAZA O CATENA GLUCIDO FOSFORICA IN CARE DEZOXIRIBONUCLEOTIDELE ALTERNEAZA CU GRUPAREA FOSFAT - PE ACEST SCHELET SE ASEAZA BAZELE AZOTATE.

FIECARE LANT POLINUCLEOTIDIC SE TERMINA CU O GRUPARE 5 FOSFAT RESPECTIV 3OH POLARITATE MOLECULEI ADN 3 5.

DNA
SugarSugar-phosphate backbone serves as a backbone. The backbone has directionality (PO4 / OH). Bases encode the genetic information.

STRUCTURA SECUNDARA A ADN

- 2 CATENE POLINUCLEOTIDICE LEGATE PRIN BAZELE AZOTATE COMPLEMENTARE: A-T si G-C. -LEGATURI PRIN PUNTI DE HIDROGEN DUBLE SAU TRIPLE. - CATENELE SUNT COMPLEMENTARE SI CODETERMINANTE.

5 PO4

3 OH

Anti-parallel Bonding

3 OH PO4 5

DOVEZI EXPERIMENTALE ALE STRUCTURII SECUNDARE A ADN

REGULA LUI CHARGAFF BAZA RATIO A+T/G+C 1

The First Clues to DNA Structure

G 22.1% 15.4% 40.4% 8.9% A 28.1% 33.6% 9.0% 42.6% T 30.1% 37.1% 11.7% 39.9% C 19.7% 13.9 38.9% 8.6%

PRINCIPIUL DENATURARII SI RENATURARII ADN

(PRIN TEMPERATURI MARI(95 GRADE CELSIUS)/ MEDIU ALCALIN MARI(95

).

Denaturation / Renaturation
The bonds that hold DNA strands together are easily broken and reformed.
5'

OH

OH

3'

3'

5'

5'

A ... T T ... A G ... C ... G ... C ... G ... C ... A ... T C ... G ... C ... G ... T ... A A ... T G ... C ... A ... T A ... T A ... T T ... A T ... A

5'

A T G G G A C C T A G A A A T T

T A C C C T G G A T C T T T A A

3'

3'

OH

OH

DOVEZI EXPERIMENTALE ALE STRUCTURII SECUNDARE A ADN IMPORTANTA: IMPORTANTA: -CAPACITATE DE AUTOREPLICARE; AUTOREPLICARE;
-CONSERVAREA INFORMATIEI GENETICE; -TRANSCRIPTIE. TRANSCRIPTIE.

- APLICATII- BIOLOGIA MOLECULARA. APLICATIIMOLECULARA.

Polymerase chain reaction (PCR)

The Big Bang

Alfred Hershey and Martha Chase (1952) DNA is genetic material. Watson and Crick (1953) DNA is a double helix.

STRUCTURA TERTIARA A ADN

FORME FIZICE ALE ADN

ADN TIP A - DEXTROGIR, SPIRE MAI APROPIATE, DEPRESIUNILE SUNT DISPUSE OBLIC REVERSIBIL CU FORMA B

ADN TIP B- APARE IN INTERFAZA- G1+S BINTERFAZAADN TIP Z- LEVOGIR, SCHELETUL GLUCIDO FOSFORIC ESTE ZNEREGULAT(ZIGNEREGULAT(ZIG- ZAG ), GUANINA ESTE LA EXTERIOR FIXEAZA RAPID SI STABIL SUBSTANTA CHIMICE CANCERIGENE

Right Handed 10.7-11 bp/turn 23A Diameter dsRNA and RNA-DNA Hybrids

From A to Z DNA
Right Handed 10-10.6 bp/turn 19A Diameter Normal DNA

Left Handed 12 bp/turn 18A Diameter dinucleotide repeats Pu-Py (GCGCGCGC)

CLASIFICAREA ADN
IN RAPORT CU STRUCTURA PRIMARA - REPETITIV - NEREPETITIV

DUPA STRUCTURA TERTIARA

DUPA TOPOGRAFIA INTRACELULARA NUCLEAR - MITOCONDRIAL

ADN NUCLEAR
CANTITATEA DE ADN NUCLEAR NU ESTE DIRECT PROPORTIONALA CU GRADUL DE EVOLUTIE A SPECIEI NU EXISTA O CORELATIE NUMAR GENE CANTITATE ADN EXPLICATIA: EXPLICATIA: GENOMUL UMAN ARE 30000 GENE CE CONTROLEAZA CARACTERE CELULARE SI INDIVIDUALE

MECANISMUL DENATURARII / RENATURARII SI HETEROGENITATEA SECVENTELOR ADN CROMOZOMAL

ADN REPETITIV
ESTE NONINFORMATIONAL INALT REPETITIV- 10-15% DIN GENOMUL CELULAR REPETITIV- 10-15% UNITATEA REPETITIVA ARE O SECVENTA DE 5-10 NUCLEOTIDE REPETATE 105-107/ POT FI MAI MULTE SECVENTE DIFERITE RENATUREAZA RAPID NU EXISTA IN CROMOZOMUL Y NU SE TRANSCRIU IN ARNm ROL DISCUTABIL IN PROTECTIE SAU ORGANIZARE

ADN REPETITIV
MODERAT REPETITIV-25-30% DIN ADN CELULAR REPETITIV-25-30% COEFICIENT DE REPETABILITATE DE 103-10-4 SECVENTA REPETITIVA 150-300 PERECHI DE 150NUCLEOTIDE INTERCALAT INTRE SECVENTELE NEREPETITIVE ARE ROL REGLATOR IN GENOMUL CELULAR LOC DE FIXARE PENTRU MOLECULELE IMPLICATE IN TRANSCRIPTIE EXISTA SECVENTE GENETIC ACTIVE- HISTOGENE, ACTIVEARNt,ARNr. ARNt,ARNr.

PALINDROMUL
SECVENTA PARTICULARA DE ADN REPETITIV ARE SIMETRIE ROTATIONALA FORMAT PE PRINCIPIUL COMPLEMENTARITATII CU BUCLE CATENARE ARE LUNGIME VARIABILA 6-12 NUCLEOTIDE STRUCTURA CU ASPECT DE AC DE PAR POATE CONTINE TRANSPOZONI ( GENE SARITOARE) IN GENOM EXISTA 120000 DE PALINDROAME ROLROL- RECUNOASTE ENZIMELE IMPLICATE IN REPLICAREA ADN SAU IN TRANSCRIPTIE ESTE RECUNOSCUT DE ENZIMELE DE RESTRICTIE SE INTILNESTE LA NIVELUL TELOMERELOR. TELOMERELOR.

ADN NEREPETITIV
SUNT SECVENTE UNICE REPREZINTA 50-70% DIN GENOM 50-70% ALTERNEAZA CU CEL NEREPETITIV DIN CROMOZOM ESTE INFORMATIONAL

GENOMUL MITOCONDRIAL

REPREZINTA 1-2% DIN TOTALUL ADN CELULAR ESTE BICATENAR, IN DUBLU HELIX, CIRCULAR POATE SUFERI MUTATII CODIFICA 30 DE GENE STRUCTURALE DETINE 17000 DE PERECHI DE BAZE GENE IMPLICATE IN LANTUL RESPIRATORRESPIRATORCITOCROM b CITOCROM-oxidaza, ATP-aza, 22 GENE CITOCROMATPPENTRU ARNt SPECIFIC SI ARNr SINTEZA ACESTOR PROTEINE POATE FI INHIBATA MEDICAMENTOS NU ARE SECVENTE NONINFORMATIONAL

GENOMUL MITOCONDRIAL
SE ASEAMANA CU ADN BACTERIAN SE REPLICA SEMICONSERVATIV INDEPENDENT DE ADN CROMOZOMIAL AVIND COMPLEX ENZIMATIC PROPRIU PENTRU REPLICARE SI TRANSCRIERE

DETERMINA EREDITATEA MATROCLINA POATE DETERMINA EXPRESIA FENOTIPICA A UNOR CARACTERE TRANZITORII SAU PERMANENTE CARACTERELE SUNT DETERMINATE DE PLASMAGENE A CAROR TOTALITATE = PLASMOM MITOCONDRIAL

CRITERII DE IDENTIFICARE A CARACTERELOR MITOCONDRIALE: REZULTATE DIFERITE DUPA FECUNDATIE CE NU RESPECTA REGULILE DETECTAREA FACTORILOR EXTRANUCLEARI CE NU RESPECTA SEGREGAREA DACA SE PRACTICA CONSANGHINIZAREA APAR CARACTERE MATERNE DUPA 2-3 GENERATII

Human Genome Project

Human Genome Project

Goals:
identify all the approximate 30,000 genes in human DNA, determine the sequences of the 3 billion chemical base pairs that make up human DNA, store this information in databases, improve tools for data analysis, transfer related technologies to the private sector, and address the ethical, legal, and social issues (ELSI) that may arise from the project.

Milestones:
1990: Project initiated as joint effort of U.S. Department of Energy and the National Institutes of Health June 2000: Completion of a working draft of the entire human genome February 2001: Analyses of the working draft are published April 2003: HGP sequencing is completed and Project is declared finished two years ahead of schedule
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

What does the draft human genome sequence tell us?

By the Numbers The human genome contains 3 billion chemical nucleotide bases (A, C, T, and G).
The average gene consists of 3000 bases, but sizes vary greatly, with the largest known human gene being dystrophin at 2.4 million bases. The total number of genes is estimated at around 30,000--much lower than previous estimates of 80,000 to 140,000. Almost all (99.9%) nucleotide bases are exactly the same in all people. The functions are unknown for over 50% of discovered genes.

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

What does the draft human genome sequence tell us?

The Wheat from the Chaff Less than 2% of the genome codes for proteins proteins.
Repeated sequences that do not code for proteins ("junk DNA") make up at least 50% of the human genome. Repetitive sequences are thought to have no direct functions, but they shed light on chromosome structure and dynamics. Over time, these repeats reshape the genome by rearranging it, creating entirely new genes, and modifying and reshuffling existing genes. The human genome has a much greater portion (50%) of repeat sequences than the mustard weed (11%), the worm (7%), and the fly (3%).

How does the human genome stack up?

Organism

Genome Size (Bases)

3 billion
2.6 billion 100 million 97 million 137 million 12.1 million 4.6 million 9700

Estimated Genes
30,000
30,000 25,000 19,000 13,000 6,000 3,200 9

Human (Homo sapiens) (Homo sapiens)

Laboratory mouse (M. musculus) Mustard weed (A. thaliana) (A. thaliana) Roundworm (C. elegans) (C. elegans) Fruit fly (D. melanogaster) (D. melanogaster) Yeast (S. cerevisiae) (S. cerevisiae) Bacterium (E. coli) (E. coli) Human immunodeficiency virus (HIV)

Future Challenges: What We Still Dont Know

Gene number, exact locations, and functions
Gene regulation DNA sequence organization Chromosomal structure and organization Noncoding DNA types, amount, distribution, information content, and functions Coordination of gene expression, protein synthesis, and post-translational events Interaction of proteins in complex molecular machines Proteomes (total protein content and function) in organisms Correlation of SNPs (single-base DNA variations among individuals) with health and disease Disease-susceptibility prediction based on gene sequence variation Genes involved in complex traits and multigene diseases Developmental genetics, genomics
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

Anticipated Benefits of Genome Research

Molecular Medicine
improve diagnosis of disease detect genetic predispositions to disease create drugs based on molecular information use gene therapy and control systems as drugs design custom drugs (pharmacogenomics) based on individual genetic profiles

Microbial Genomics
rapidly detect and treat pathogens (disease-causing microbes) in clinical practice develop new energy sources (biofuels) monitor environments to detect pollutants protect citizenry from biological and chemical warfare clean up toxic waste safely and efficiently
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

Anticipated Benefits of Genome Research-cont. ResearchRisk Assessment

evaluate the health risks faced by individuals who may be exposed to radiation (including low levels in industrial areas) and to cancer-causing chemicals and toxins

Bioarchaeology, Anthropology, Evolution, and Human Migration

study evolution through germline mutations in lineages study migration of different population groups based on maternal inheritance study mutations on the Y chromosome to trace lineage and migration of males compare breakpoints in the evolution of mutations with ages of populations and historical events

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

Anticipated Benefits of Genome Research-cont. ResearchDNA Identification (Forensics)

identify potential suspects whose DNA may match evidence left at crime scenes exonerate persons wrongly accused of crimes identify crime and catastrophe victims establish paternity and other family relationships identify endangered and protected species as an aid to wildlife officials (could be used for prosecuting poachers) detect bacteria and other organisms that may pollute air, water, soil, and food match organ donors with recipients in transplant programs authenticate consumables such as caviar and wine

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

Medicine and the New Genetics

Gene Testing Pharmacogenomics Gene Therapy

Anticipated Benefits:
improved diagnosis of disease earlier detection of genetic predispositions to disease rational drug design gene therapy and control systems for drugs personalized, custom drugs

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

ELSI: Ethical, Legal, and Social Issues

Privacy and confidentiality of genetic information. Fairness in the use of genetic information by insurers, employers, courts, schools, adoption agencies, and the military, among others. Psychological impact, stigmatization, and discrimination due to an individuals genetic differences. Reproductive issues including adequate and informed consent and use of genetic information in reproductive decision making.

Clinical issues including the education of doctors and other health-service providers,
people identified with genetic conditions, and the general public about capabilities, limitations, and social risks; and implementation of standards and quality-control measures.
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

ELSI Issues (cont.)

Uncertainties associated with gene tests for susceptibilities and complex conditions (e.g., heart disease, diabetes, and Alzheimers disease). Fairness in access to advanced genomic technologies. Conceptual and philosophical implications regarding human responsibility, free will vs genetic determinism, and concepts of health and disease. Health and environmental issues concerning genetically modified (GM) foods and microbes. Commercialization of products including property rights (patents, copyrights, and trade secrets) and accessibility of data and materials.

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

HapMap
An NIH program to chart genetic variation within the human genome

Begun in 2002, the project is a 3-year effort to construct a map of the patterns of SNPs (single nucleotide polymorphisms) that occur across populations in Africa, Asia, and the United States. Consortium of researchers from six countries Researchers hope that dramatically decreasing the number of individual SNPs to be scanned will provide a shortcut for identifying the DNA regions associated with common complex diseases Map may also be useful in understanding how genetic variation contributes to responses in environmental factors