摘要
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。
Aim To isolate and express the urease B (ureB)gene from Helicobacter pylori.Methods The ureB gene was amplified from H.pylori chromosomal DNA by PCR.The ureB gene was recombined in vitro with expression vector pQE30and was transformed into E.coli M15cells.Results The 1.7kb ureB gene was successfully isolated.Recom-binant E.coli strains expressed UreB were obtained,the expression protein amounted to 34.0%of the tota l bacterial protein after induced wi th IPTG for 4h at 37℃.Conclusion Cloning and high -expression of ureB gene from H.lay the foundation for co nstructing the H.pylori vaccine.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第5期464-465,共2页
Chinese Journal of Cellular and Molecular Immunology
基金
全军重点课题基金资助
No.98Z071
