PCR直接测序方法及其在肿瘤研究中的应用

The Method of PCR Direct Sequencing and It'sApplication in Cancer Research.

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￣（－３５）ＳｄＡＴＰ直接掺入，ＴａｑＤＮＡ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 com-bined PCR amplification with nucleic acid se-quencing technique. According to this tech-nique, direct sequencing DNA strand of PCRamplification using PCR primer, a-￣(35) S dATPand Taq DNA polymerase. The experimentshowed that it is simple, rapid and stable.This method was used to analvze the tumorsuppressor gene p53 mutation in humanesophageal eancer. It was found that therewere point mutation.insertion and deletionframeshift mutation of p53 gene in humanesophageal cancer. Intron 5 and 8 sequences of p53 gene in human and Rhesus monkey weresequenced and in monkey they are 81 and 92nucleotides respectively.