结核分枝杆菌基因组学精析.ppt

结核分枝杆菌基因组学 黄海荣 国家结核病参比实验室  提纲 一、结核分枝杆菌基因组总论 目前一些相关实验室技术的遗传学背景 1基因分型技术 2耐药的分子生物学诊断 3新近的新疫苗、免疫诊断技术  Genome ofM. tuberculosis More Than Maps of 4,000 genes other spp. nearly identical M. tuberculosis 4,411.529bp Cole et al.(1998) Nature393:537-544  A diamete Large groove (-2A 34 Length of one Small groove pposite polarity Axis  The Structure of DNA 5′end bases C minor groove 3end p-O 0.34nm major g roove AG A 0-P-O O=P-O hydrogen bond phosphodiester 5′end Figure 4-5. Molecular Biology of the Cell, 4th Edition  TABLE 11.1 The genetic Code First Third Position econd Positio (5-end) Positio (3’end) UUU Phe UCU Ser UAU Tyr UGU CYS UUC Phe UCC Se UAC TVI UGC C\ UUA Leu UCA Ser UAA Stop UGA Stop UUG Leu UCG Ser UAG Stop UGG Trp G CUU Leu CCU Pro CAU His CGU Arg CUC Lo CCC Pro CAC His CGC Arg CUA Let CCA Pro CAA GIn CGA Arg CUG Leu CCG Pro CAG GIn CGG Arg AUU lle ACU Thr AAU Asn AGU Ser AUC lle ACC Thi AAC Asn AGC Ser A AUA le ACA Thr AAA I AGA Arg AUG Meta ACG Thr AAG Lv AGG Arg UCAGUCAGUCAGUCA GUU Val GCU Ala GAU Asp GGU Gly GUC Val GCC Ala GAC Asp GGC Gly GUA Val GCA Ala GAA Glu GGA GI GUG Val GCG Ala GAG Glu GGG Gly  Native double Strands Double helix strands reformed Heat M Cool Denaturation Renaturation  Hierarchical Shotgun Sequencing Method 这 Genomic dNA BAC Library Create contig mi Sequence each conti with Shotgun Appoach Align Contiguous Sequences GCHTICGAGILHCCHGACRACO GG TACTGAGGACCCRAGAGC F.rj  Whole genome shotgun sequencing of shotgun ic DNA partial digestion overlapping DNA 了N f 1-2 kb  Whole genome Shotgun Sequencing Method Genomic dna Sequence Each Fiagment with Shotgun Approach CCATVICGAGTIACCTGGACRACCAGT lign Contiguous Sequences TSP. Gtnelate Fmished secnutct