基因诊断常用技术

1、l1950s-Rosalind Franklin (1920-1957) discover chemical structure of DNA, starting a new branch of science-molecular biology.lWatson and Crick made a model of the DNA molecule and proved that genes determine heredity. l1957-Arthur Kornberg (1918- ) of the U.S. produced DNA in a test tube. l1966-The G

2、enetic code was discovered; scientists are now able to predict characteristics by studying DNA. This leads to genetic engineering, genetic counseling. l1972-Paul Berg (1926- ) of the U.S. produced the first recombinant DNA molecule. l1983-Dr. Kary Mullis discovered the PCR procedure, for which he wa

3、s awarded the Nobel prize. l2000-J. Craig Ventor, along with Francis Collins, jointly announce the sequencing of the entire human genome Enpromoter5E1GTAG15UT33UT人类结构基因包括人类结构基因包括4个区域：编码区：包括外显子、内含子个区域：编码区：包括外显子、内含子 前导区：编码区上游前导区：编码区上游 尾部区：编码区下游尾部区：编码区下游 调控区：包括启动子、增强子等调控区：包括启动子、增强子等E2I2转录起始点转录起始点终止密码终止

4、密码AATAAA转录终止点转录终止点 1 ggggacagcc agggacaggc agacatgcag ccagggctcc agggcctgga caggggctgc 61 caggccctgt gacaggagga ccccgagccc ccggcccggg gaggggccat ggtgctgcct 121 gtccaacatg tcagccgagg tgcggctgag gcggctccag cagctggtgt tggacccggg 181 cttcctgggg ctggagcccc tgctcgacct tctcctgggc gtccaccagg agctgggcgc 241

5、ctccgaactg gcccaggaca agtacgtggc cgacttcttg cagtgggcgg agcccatcgt 第一代诊断技术第一代诊断技术- - 限制性片段长度多态性检测（限制性片段长度多态性检测（RFLPRFLP）镰性细胞贫血镰性细胞贫血第一代诊断技术第一代诊断技术- - Southern Southern杂交杂交SouthernSouthern杂交基本步骤杂交基本步骤Deletion第一代诊断技术第一代诊断技术- - DNA DNA体外扩增技术体外扩增技术（PCR）PCRPCR技术出现的背景技术出现的背景 1955年发现 DNA polymerase I； 70年代的初期发现具有实验价值及可得性Klenow fragment 1971年Dr. Kjell Kleppe 提出类似基因修复复制 的PCR原始雏形概念； 1976年从热泉的细菌(Thermus Aquaticus) 中分离出能耐高温 Taq polymerase； 1983年Dr