Construction and eukaryotic expression of recombinant plasmid pEGFP-N1-SrV+encoding the variable region of the surface protein of Streptococcus mutans
10.3760/cma.j.issn.0254-5101.2011.04.009
- VernacularTitle:变形链球菌表面蛋白可变区真核载体质粒pEGFP-N1一SrV+的构建及在293T细胞的表达
- Author:
Kuifang HE
;
Xiaoguang YU
;
Yihuai PAN
;
Jianguo LIU
;
Qingguo QI
- Publication Type:Journal Article
- Keywords:
Streptococcus mutans;
Variable region of surface proteins;
Eukaryotie expression vector plasmid pEGFP-N1;
293T cells
- From:
Chinese Journal of Microbiology and Immunology
2011;31(4):324-329
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant plasmid pEGFP-N1-SrV+ and evaluate the expression of SrV+in mammalian 293T cells.nethods srv+.a gene encoding the vailable region of the surface protein of the Streptococcus mutans OMZ175.was cloned chemically based on its reported nucleotide sequence.The eukaryotic expression plasmid,pEGFP-N1-SrV+,was constructed by introducing the srv+ gene into the Kpn Ⅰ/Xho Ⅰ site of pEGFP-NI.The recombinant plasmid pEGFP-N1-SrV+was transfected into 293T cells with lipofectamine and the expression level of SrV+was evaluated.Results The eukaryotic expression plasmid pEGFP-N1-SrV+was constructed successfully.GFP was observed by green fluorescent microscope.and a 72 × 1 03 protein was detected bv Westem blot.Real-time RT-PCR analysis revealed that the expression of the pEGFP-N1-SrV+in 293T was excellent and significant compared the control group. Conclusion The recombinant plasmid pEGFP-N1-SrV+was successfully constructed.which could encode the expression of SrV+after transfected into the mammalian 293T Cells.
