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Human genome project
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- 2. What is Genome? SalientFeatures Of Human Genome Participants of the HGP Goals DNA Sequencing Application Human Genome Project Issue of concern Future Challenges: what we still don’t know
- 3. A genome isan organism’s complete set of DNA, including all of its genes. Each genome contains all of the information needed to build and maintain that organism. The genome includes both the genes and the non-coding sequence of DNA/RNA.
- 4. 1 bp =660 Dalton Genome of organism differ from others Arrangement of bases Number of base pairs 1000 bp = 1 kb 1000 kb = 1 Mb ( 10 6 bp) 1000 Mb = 1 Gb (10 9 bp)
- 5. Erwin Chragaff Organism AT G C A/T G/C E.Coli 26 23.9 24.9 25.2 1.09 0.99 Yeast 31.3 32.9 18.7 17.1 0.95 1.09 Drosophila 27.3 27.6 22.5 22.5 0.99 1.00 Honey Bee 34.4 33.0 16.2 16.4 1.04 0.99 Mouse 29.2 29.4 21.7 19.7 0.99 1.10 Human(liver) 30.7 31.2 19.3 18.8 0.98 1.03 Chargaff’s rule G=C A=T DNA composition varies from species to species
- 6. SPECIES GENOME SIZE(Mb) No. of Chromosome(n) FUNGI SACCHAROMYCES CEREVISIAE 12.1 16 ASPERGILLUS NIDULANS 25.4 8 PLANTS ARABIDOPSIS THALIANA 125 5 ORYZA SATIVA 430 12 ZEA MAYS 2500 10 PISUM SATIVUM 4800 14 TRITICUM AESTIVUM 16000 24 FRITILLARIAASSYRIACA 120000 12 INVERTEBRATES CAENORHABDITIS ELEGANS 97 7 DROSOPHILA MELANOGASTER 180 4 BOMBYX MORI 490 28 VERTEBRATES MUS MUSCULUS 3300 19+X & Y HOMO SAPIENS 3200 22 + X & Y
- 7. GENE NUMBER SIZE(bp) VIROIDS PSTV RNA 0 359 SS RNA VIRUSES STNV 1 1300 PHAGE MS2 4 3569 TMV 4 6400 INFLUENZA 12 13500 DS RNA VIRUSES REOVIRUS 22 23000 SS DNA VIRUSES PHAGE x174 11 5387 DS DNA VIRUSES SV 40 6 5226 PHAGE T4 200 165000 ORGANISMS MYCOPLASM 500 <106 BACTERIA 2-4000 <107 FUNGI 6000 1.3 X 107 FLIES 12000 1.6 X 108 MAMMALS 20000 3 X 108 PLANTS 50,000 < 1011
- 8. ATCGCGCTA AAATGGTCA TATTAGGC TACGGTTA CGATGGCT A BOOK 1000PAGES E.COLI 300 PAGES YEAST 1 BOOK DOSOPHILA 10 BOOKS HUMAN 200 BOOKS EACH PAGE 50 LINES EACH LINE 100 LETTERS ATCGCGCTA AAATGGTCA TATTAGGC TACGGTTA CGATGGCT
- 9. Human genome consistsinformation of 23 chromosome (22Autosomes +1X chromosome) Human genome contains over 3 billion nucleotide pair, about 30,000 genes & 3000 bases. Only 3% of genome codes for amino acid sequence of polypeptide . Only 0.2% of base pair is different in individual.
- 10. Structure of chromosome1 (Has largest genes i.e., 2968genes) Y chromosome has lowest genes(i.e., 231)
- 12. National Institute OfHealth (NIH) The Department Of Energy (DOE) USA UK FRANCE GERMANY JAPAN CHINA Total expenditure $3/bp ( 9billion US $) Time Period= 13 years Celera: 5 individuals; HGSC: ‘many’
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- 15. Maxam-Gilbert chemical sequencing Sangerchain termination sequencing Automated Sequencing The term DNA sequencing encompasses biochemical methods for determining the order of the nucleotide bases, Adenine, Guanine, Cytosine & Thymine in DNA oligonucleotide.
- 16. 32 P 5’ 5’ 3’ 3’ 5’ 3’ TAGTCGCAGGAC ds DNA ssDNA Radio labelled G A+G T+C C 32PTAG 32PTA 32PT 32PTAGTC 32PTAGTCG 32PTAG 32PTAGT 32PTAGTCGC 32PTAGTCGCAG 32PTAGTCG 32PTAGTC 32PTAGTCGCAGGAC 32PTAGTCGCAGG 32PTAGTCGCA 32PTAGTCGC 32PTAGTCGCAG 32PTAGTCGCAGGAC 32PTAGTCGCAGG 32PTAGTCGCAGGC
- 17. G A+G T+CC Bands on Autoradiogram 3’ C A G G A C G C T G A T 5’ Sequencing gels are read from bottom to top (5′ to 3′). 5’ G T C C T G C G A C T A 3’ Sequence of base on the Complementary strand. Sequence of base on the analysed strand. ELECTROPHORESIS
- 18. ssDNA , LabelledPrimer , DNA Polymerase ,Excess dNTPs 3’ 3’ 5’ 5’ 5’ 3’ TAGTCGCAGGACCG ds DNA ss DNA A C G T dd ATP dd CTP dd TTP dd GTP ATCAGCGTCCTGGC ATCAGCGTCCTGGC ddATP ATCddATP ATddCTP ATCAGddCT ATCAGCGTddCTP ATCAGCGTCddCTP ATCAGCGTCCTGGddCTP ATCAddGTP ATCAGCddGTP ATCAGCGTCCTddGTP ATCAGCGTCCTGddGTP AddTTP ATCAGCGddTTP ATCAGCGTCCddTTP P 32
- 19. Sequencing gels are read frombottom to top (5′ to 3′). A C G T 3’ C G G T C C T G C G A C T A 5’ 5’ G C C A G G A C G C T G A T 3’ Sequence of base on the Complementary strand. Sequence of base on the analysed strand. Bands on Autoradiogram ELECTROPHORESIS
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- 22. Fox2B Dysbindin-1 CHRM-2 Williams syndrome WILLIAMS SYNDROME; LOWIN GENERAL INTELLIGENCE BUT GOOD IN LANGUAGE : CHROMOSOME 11 SPECIFIC LANGUAGE IMPAIRMENT : LOW LINGUISTIC ABILITY WITHOUT AFFECTING INTELLIGENCE : CHROMOSOME 7
- 26. Privacy & confidentialityof genetic information. Psychological impact, stigmatization & discrimination. Uncertainties associated with gene test for susceptibilities and complex conditions. Clinical issues. Commercialization of products. Fairness in access to advanced genomic technologies . Health & environmental issues .
- 27. Gene number, exactlocation, function & regulation Chromosomal structure & organisation. Non coding DNA types, amount, distribution, information content, & function. Disease susceptibility prediction based on gene sequence variation. Genes involved in complex traits & multigene disease. Developemental genetics & genomics.