Construction and expression of recombinant eukaryotic plasmid pVAX1-SPG
WANG Yi-dan
SUN Tian-tong
LIU Jian-guo
CHAI Qiao-xue
QU Yun-peng
YU Xiao-guang
BAI Guo-hui
Tian Yuan
Han Qi
Abstract：AIM::To construct the eukaryotic plasmid including surface protein antigen ( SpaP) P domain (SpaP-P) and glucan binding domain (GBD) of glucan binding protein A (GbpA) of Streptococcus mutans, and to observe the expression of recombinant plasmid pVAX1-SPG in mammalian cells 293T. METHODS: The eukaryotic plasmid pVAX1-SPG carrying encoding gene of GBD of GbpA and SpaP-P of SpaP of Streptococcus mutans was con-structed by recombinant DNA technology. The eukaryotic plasmid was transfected into 293T cells by lipofectamine rea-gent. The transient expression of the protein in 293T cells was detected by immunochemistry technique and Western-blot. RESULTS:The recombinant plasmidp VAX1-SPG was obtained and identified by restriction endonuclease anal-ysis. Immunohistological staining showed that positive expression of SpaP/P-GBD was detected in the plasma of 293T cells transfected with recombinant plasmid pVAX1-SPG. Western-blot confirmed the expression of the fusion protein SpaP/P-GBD in 293T cells. CONCLUSION:The eukaryotic plasmid pVAX1-SPG can be constructed. The target protein SpaP/P-GBD can be expressed correctively in 293T cells.
Keywords：streptococcus mutanssurfaceprotein antigenglucan-binding proteinmammalian cellseukaryo-ticexpression
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 645-650 )