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目的构建HP450基因的克隆载体。方法用逆转录聚合酶链反应(RT-PCR)技术克隆HP450基因,T-A连接到pMD18-T载体。以菌液为模板做PCR、酶切和测序鉴定pMD18-HP450重组质粒载体。结果菌液PCR扩增出目的条带、重组体的酶切产物合理,测序显示目的基因与基因库中的H1基因100%同源。结论实现了HP450基因的克隆及其重组质粒载体pMD18-HP450的构建。
Objective To construct cloning vector of HP450 gene. Methods HP450 gene was cloned by reverse transcription polymerase chain reaction (RT-PCR) and T-A was ligated into pMD18-T vector. The bacterial liquid as a template to do PCR, digestion and sequencing identified pMD18-HP450 recombinant plasmid vector. Results The bacterial bands were amplified by PCR, and the products of restriction endonuclease digestion were reasonable. The sequencing showed that the target gene was 100% homologous to the H1 gene in the gene pool. Conclusion The cloning of HP450 gene and the construction of recombinant plasmid vector pMD18-HP450 were achieved.