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目的克隆并表达幽门螺杆菌尿素酶B基因。方法提取幽门螺杆菌染色体DNA用PCR方法扩增尿素酶B基因。将其克隆至表达载体pQE30,转化大肠杆菌M15,IPTG诱导表达。结果分离得到了1.7kb的ureB基因片段,并在大肠杆菌中实现了该基因的高效表达。在37℃诱导表达4h后,表达产物约占细菌总蛋白的34.0%。表达以可溶性蛋白和包涵体两种形式存在,其中主要是包涵体的形式,目的蛋白占不溶性蛋白的55.8%。结论幽门螺杆菌ureB基因的克隆与表达为Hp疫苗的研制打下了基础。
Objective To clone and express Helicobacter pylori urease B gene. Methods Chromosomal DNA of Helicobacter pylori was extracted for PCR amplification of urease B gene. It was cloned into the expression vector pQE30, transformed into E. coli M15, IPTG induced expression. Results A 1.7 kb ureB gene fragment was isolated and highly expressed in Escherichia coli. After induction for 4h at 37 ℃, the expression product accounted for about 34.0% of total bacterial protein. The expression of soluble protein and inclusion bodies exist in two forms, mainly in the form of inclusion bodies, the target protein accounted for 55.8% of insoluble protein. Conclusion Cloning and expression of ureB gene of Helicobacter pylori lay the foundation for the development of Hp vaccine.