摘要
目的分段表达幽门螺杆菌(HP)尿素酶B亚单位(UreB),并对其进行免疫学性质分析,为精确定位其保护性表位奠定基础。方法用生物信息学软件分析UreB全长基因,选择分段点,PCR分别扩增其5个片段,构建于pET-11c(+)原核表达载体,并转化大肠杆菌BL21(DE3),用IPTG诱导表达,SDS-PAGE分析蛋白表达情况,ELISA及Western blot分别鉴定其免疫原性及抗原性。结果成功克隆了UreB的5个基因片段,基因测序结果与Genbank公布的序列一致.经SDS-PAGE分析,5个蛋白表达相对分子质量大小都与预期相符,在大肠杆菌中实现了表达。ELISA和Western blot分析显示,表达的5个蛋白表现出良好的抗原性和免疫原性。结论 UreB的分段表达为进一步研究UreB保护性表位及HP疫苗鉴定奠定了新的基础。
With consideration of making a foundation for further investigation on protective epitopes of urease B （UreB） subunit, we express the recombinant gene fragments of UreB subunit of Helicobacter pylori in E. coil and analyze its immunological characteristics. We analyzed the protein structure of UreB by the DNASTAR software, and then amplified 5 UreB fragments by polymerase chain reaction, whose nucleotide sequences were consistent with the information in Genbank. The enzyme-digested target fragments was cloned into PET-11c （＋） expression vector, and transferred into E. coli BL21 （DE3）. After induction with IPTG, SDSPAGE confirmed all the 5 fragments of UreB could express in E. coli BL21 （DE3）. ELISA and Western blotting showed all of the recombinant proteins had good antigenicity and immunogenicity to specific antibodies. The expression of UreB fragments made a new foundation for further investigation on protective epitopes of UreB and vaccine against HP.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第3期260-263,共4页
Immunological Journal
基金
国家自然科学基金课题(30771992)
"十一五"重大专项课题(2008ZX10004-015)
