南方医科大学学报2012，Vol.32Issue(2)：185-188,4.DOI:44-1627/R.20120209.1413.018
人Tau基因多表位肽段原核表达载体的构建与表达
Construction of a prokaryotic expression vector of human tau multi-epitope peptide and immunogenicity of the expressed product
摘要
Abstract
Objective To construct a prokaryotic expression vector of human tau multiepitope peptide for examining the immunogenicity of a TauPl/P2 DNA vaccine in mice using the expressed product. Methods The coding sequence of Tau multiepitope peptide gene was amplified from the plasmid pVAXl-Tau by PCR and inserted into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant plasmid pGEX-4T-2-TauPl/P2. The positive recombinants were transformed into E.coli BL21 cells, and the expression of fusion protein GST-TauPl/P2 was induced by IPTG and identified by SDS-PAGE. Mice was immunized with TauPl/P2 DNA vaccine and the production of the specific antibodies was detected by Dot-blot analysis using the purified fusion protein. Results A gene fragment 300 bp in length was amplified. Enzyme digestion and DNA sequencing verified correct construction of the prokaryotic expression plasmid pGEX-4T-2-TauPl/P2. The expression of target fusion protein GST-TauPl/P2 was detected by SDS-PAGE. Specific antibodies against TauPl/P2 were detected in the serum of mice immunized with the DNA vaccine using GST-TauPl/P2 fusion protein. Conclusion The constructed prokaryotic expression plasmid of human Tau multiepitope peptide is capable of expressing the target fusion protein, which specifically recognizes the specific antibodies against TauPl/P2 in mice immunized with TauPl/P2 DNA vaccine.关键词
Tau/质粒/原核表达/融合蛋白Key words
TauPl/P2/ plasmid/ prokaryotic expression/ fusion protein分类
生物科学引用本文复制引用
孙海涛,杨化强,杨璐军,李正阳,杜谋选,陈宇新,姜晓丹..人Tau基因多表位肽段原核表达载体的构建与表达[J].南方医科大学学报,2012,32(2):185-188,4.基金项目
国家自然科学基金(81171179) （81171179）
广东省及广州市科技计划重点项目[粤财企(2008) 258-2008A030201019,穗科字(2008) 3-2008A1.E4011-6,09B52120112-2009J1-C418-2] （2008）
