HP450基因的克隆及其重组载体pMD18-HP450的构建与鉴定

HP450 Gene Clone and Construction and Identification of the Recombinant Vector pMD18-HP450

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 The HP450 gene was obtained by reverse transeriptase-polymerase chain reaction （RT-PCR）. It was cloned into pMD18-T vector using T-A clone methed. We identified the recombinant plasmidvector pMD18-HP450 by restriction digestion, sequencing and PCR using bacteria as the templet. Results The aim strip appeared after the PCR and the restriction digestion production were logical. The sequencing showed that 100% nucleotides were homologous between aim gene and Hlgene in gene bank. Conclusion The HP450 gene had been cloned. The recombinant vector pMD18-HP450 is constructed correctly.