PCR直接测序技术是PCR扩增与核酸测序技术相结合的一种方法.根据此技术的原理,建立了一种以PCR扩增引物为测序引物,α-35S dATP直接掺入,Taq DNA聚合酶直接测序PCR扩增产物的方法.实验表明:该方法简便、快速、稳定.用此方法对人食管癌组织中的抗癌基因p53进行了突变测序分析,发现食管癌组织中p53存在点突变,插入、丢失移码突变.并用此方法对人和恒河猴的p53内含子序列进行了测定,发现猴第5内含子为81个核苷酸,第8内含子为92个核苷酸.
PCR direct sequencing is a method which combined PCR amplification with nucleic acid sequencing technique. According to this technique, direct sequencing DNA strand of PCR amplification using PCR primer, α-35 S dATP and Taq DNA polymerase. The experiment showed that it is simple, rapid and stable. This method was used to analvze the tumor suppressor gene p53 mutation in human esophageal eancer. It was found that there were point mutation,insertion and deletion frameshift mutation of p53 gene in human esophageal cancer. Intron 5 and 8 sequences of p53 gene in human and Rhesus monkey were sequenced and in monkey they are 81 and 92 nucleotides respectively.
李华川,陆士新. PCR直接测序方法及其在肿瘤研究中的应用[J].生物化学与生物物理进展,1994,21(2):167-170
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