幽门螺杆菌尿素酶基因ureB植物双元表达载体的构建

摘要

目的:构建含有人幽门螺杆菌(H.pylori,Hp)ureB基因的植物表达载体.方法:采用高保真PCR技术从质粒pMED-ureB中扩增出ureB基因,构建质粒pUC18-ureB,再将pUC18-ureB酶切,将ureB基因与质粒pBI121连接.结果:构建的PBI121-ureB经PCR、酶切鉴定和测序分析证明,插入的基因片段为ureB基因,长度为1 716 bp,与GenBank报道的核苷酸序列同源性为99.76%,氨基酸序列同源性为100%.结论:成功构建了ureB基因的植物表达载体.%Aim:To construct plant expression vector for Helicobacler pylori( HP) gene ureB. Methods:Gene ureB was amplified by high-fidelity PCR from plasmid pMED19-ureB and inserted into the corresponding endonuclease enzyme digested plasmid pUC18. The recombinant plasmid pVClS-ureB was digested and ureB gene was inserted into the plasmid pBI121. The recombinant vector pBI121-ureB was identified by PCR and restricted endonuclease enzyme. Results: Enzyme digestion analysis and sequencing results showed that the target gene was 1 716 bp and had been inserted into recombinant vector. Compared with gene reported by CenBank, the target gene had 99.76% homology in uncleotide acid sequence and 100% homology in amino acid sequenc. Conclusion: The plant expression vector of ureB has been constructed successfully.