摘要
目的通过自诱导表达系统重组表达结核分枝杆菌H37Rv菌株可溶性蛋白抗原Mtb8.1,并对其抗原性进行分析。方法应用PCR技术扩增结核分枝杆菌H37Rv菌株Mtb81蛋白抗原编码序列,将其插入克隆载体pMD18-T,酶切后插入表达载体PET28a,通过优化培养基和发酵工艺,摸索出一条高效的可溶表达方法,以间接ELISA法研究其免疫学特性。结果与结论重组的Mtb81高效可溶性表达,纯化后纯度达95%以上;用纯化的重组蛋白抗原作为包被抗原建立的间接ELISA法检测200份血清样本,阳性率达34.0%,阴性率达91.0%,,符合率达62.5%。
In this study,we recombinantly expressed soluble protein Mtb8.1 antigens of Mycobacterium tuberculosis H37Rv strain by self-inducible expression system and analyzed its antigenicity.Mtb8.1 antigen coding sequence of Mycobacterium tuberculosis H37Rv strain was amplified and inserted into the plasmid pMD18-T.After that,the new recombinant was in-serted into an expression vector PET28a after digestion.An efficient method for soluble expression was worked out by optimizing the culture medium and fermentation process.The immunological characteristics of the new recombinant protein Mtb8.1 was studied by indirect ELISA.Results indicated that the recombinant Mtb8.1 showed efficiently soluble expression,and the purity of rMtb8.1 was over 95% after purified.A total of 200 serum samples were detected by indirect ELISA assay with the purified rMtb8.1 as envelope antigen,and the positive rate,negative rate and coincidence were 34.0%,91.0% and 62.5%,respectively.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2014年第5期535-537,共3页
Chinese Journal of Zoonoses
基金
国家自然科学基金(No.81260456)资助~~
