[Construction of eukaryotic expression vector containing foot-and-mouth disease virus VP1 gene and its expression in dendritic cells].
OBJECTIVE
To construct the eukaryotic expression vectors for the FMDV VP1 gene and transfect dendritic cells(DC) for the expression of VP1.


METHODS
The plasmid pMD-18-VP1 was subcloned into pcDNA3.1(+) to construct recombinant eukaryotic expression plasmids named as pcDNA3.1-VP1. The recombinant plasmids were transfected into DC by lipofectamine method and the positive cell clones were screened with G418. The expression of FMDV VP1 gene in DC was determined by Western blot.


RESULTS
The correct construction of pcDNA3.1-VP1 was identified by means of restriction enzyme analysis and nucleotide sequence determination. It was showed by SDS-PAGE and Western blot that transfected cells, DC, expressed FMDV VP1 gene constantly.


CONCLUSIONS
The pcDNA3.1-VP1 eukaryotic expression plasmids are successfully constructed and VP1 gene can be sustainly expressed in DC.
Li
Jie
J
Department of Immunology and Cell Biology, College of Veterinary Medicine, Agricultural University of Hebei, Baoding 071000, China. lijie2006698@yahoo.com.cn
Wang
Ruo
R
Shi
Wei
W
Bian
Hai-xia
HX
Zhang
Li
L
Zhang
Lei
L
Wang
Jia-xin
JX
D000818:Animals; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology*; D005536:Foot-and-Mouth Disease / Q000235:genetics / Q000276:immunology / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D036022:Capsid Proteins; C442501:VP1 protein, Foot-and-mouth disease virus