Details

Autor(en) / Beteiligte

• Kong, Fan-zhi
• Zheng, Dong
• Su, Zhao-liang
• She, Peng
• Xu, Xin-xin
• Tian, Sha-sha
• Zhang, Pan
• Chen, Jian-guo
• Xu, Hua-Xi

Titel

Construction of eukaryotic co-expression vector of Porphyromonas gingivalis outer membrane protein ragB and glucocorticoid-induced tumor necrosis factor receptor ligand (GITRL) and its immunogenic analysis

Ist Teil von

• Xi bao yu fen zi mian yi xue za zhi, 2012-06, Vol.28 (6), p.588

Ort / Verlag

China

Erscheinungsjahr

2012

Quelle

MEDLINE

Beschreibungen/Notizen

• To construct eukaryotic co-expression vector of Porphyromonas gingivalis outer membrane protein ragB and mouse glucocorticoid-induced tumor necrosis factor receptor ligand (mGITRL) and to analyze its immunogenicity in vivo. The ragB gene was obtained from pMD18-T-ragB, and then cloned into the eukaryotic expression vector pIRES and pIRES-mGITRL, respectively. The eukaryotic expression vectors: pIRES-ragB and pIRES-ragB-mGITRL were identified by double enzyme digestion and DNA sequencing, then transfected into COS7 cells by Lipofectamine(TM);2000. The expressions of ragB or mGITRL in COS7 cells were detected by Western blotting. The mice were immunized with the recombinant pIRES-ragB-mGITRL plasmid. The serum antibody level was determined by ELISA. pIRES-ragB and pIRES-ragB-mGITRL plasmids were successfully constructed. Western blotting showed that the targeted gene was over-expressed in COS7 cells and skeletal muscle cells, respectively. The high titers of antibodies against RagB were detected in mouse serum.