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目的:在正常人肝组织中克隆nm23 -H1cDNA基因,构建腺病毒克隆载体。方法:应用逆转录多聚酶链式反应(RT- PCR)技术,从正常人肝组织中扩增出nm23- H1cDNA,连接到质粒pMD18- T上,经测序、鉴定,与腺病毒穿梭质粒正向连接,构建腺病毒克隆载体。结果:克隆的基因经测序鉴定,与已知nm23-H1cDNA编码区基因100%符合,以此基因成功构建正向腺病毒穿梭质粒载体。结论:利用分子克隆技术,成功克隆中国人nm23- H1cDNA基因,构建出正向重组腺病毒穿梭质粒载体。
Objective: To clone the nm23-H1 cDNA gene in normal human liver tissue and construct adenoviral cloning vector. Methods: nm23-H1 cDNA was amplified from normal human liver tissue by reverse transcription-polymerase chain reaction (RT-PCR) and ligated into plasmid pMD18-T. After sequencing and identification, the plasmid was ligated with adenovirus shuttle plasmid , Construction of adenovirus cloning vector. Results: The cloned gene was identified by sequencing and was 100% identical to the known coding region of nm23-H1 cDNA. The positive shuttle plasmid vector was successfully constructed. Conclusion: Using molecular cloning technique, we successfully cloned Chinese nm23-H1 cDNA gene and constructed a shuttle plasmid vector of recombinant adenovirus.