Construction of the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila and its expression in NIH3T3 cells.
- Author:
Cai-xia JING
1
;
Jia-zhou YANG
;
Ming-jie LIU
;
Jia-nan XU
;
Wang GUAN
;
Ying XU
;
Jian-ping CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bacterial Vaccines; genetics; CpG Islands; genetics; Legionella pneumophila; genetics; Mice; NIH 3T3 Cells; Plasmids; genetics; Recombinant Proteins; biosynthesis; genetics; immunology; Transfection; Virulence Factors; biosynthesis; genetics
- From: Journal of Southern Medical University 2010;30(3):468-481
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila for its expression in NIH3T3 cells.
METHODSlvgA gene flanked with CpG motifs of Legionella pneumophila was amplified by PCR. The PCR products was inserted into the eukaryotic expression plasmid pcDNA3.1/myc-his(+) to construct the recombinant plasmid pclvgA/CpG, which was subsequently transfected into NIH3T3 cells via lipofection. Immunofluorescence analysis was carried out to detect the transient expression of the plasmid in the cells.
RESULTSSequence analysis showed that the recombinant plasmid pclvgA/CpG contained the lvgA/CpG fragment with a length of 657 bp, encoding a protein of 27.7 Ku. Immunofluorescence analysis identified the transient expression of the recombinant plasmid pclvgA/CpG in NIH3T3 cells.
CONCLUSIONThe lvgA gene flanked with CpG motifs of Legionella pneumophila has been constructed successfully, and the transient expression of the recombinant plasmid pclvgA/CpG can be detected in NIH3T3 cells.
