Construction and eukaryotic expression of recombinant plasmid encoding fusion protein of goat complement C3d and foot-and-mouth disease virus VP1 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 209-213, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-276138
ABSTRACT
We constructed a recombinant plasmid encoding VP1 gene of O type foot-and-mouth disease virus fused to a molecular adjuvant, goat complement C3d gene. The goat C3d gene was cloned and three copies were tandem-linked with the linker (G4S)2 sequence. VP1 gene of O type foot-and-mouth disease virus was linked to three tandem repeats of C3d through the linker sequence and cloned into pUC19 to obtain the recombinant plasmid pUC19-VP1-C3d3. The VP1-C3d3 fusion gene was then subcloned into the eukaryotic vector pcDNA3.1(+) that had been modified to contain the tissue plasminogen activator (tPA) leader sequence to obtain pcDNA3.1-tPA-VP1-C3d3. HeLa cells were transfected with pcDNA3.1-tPA-VP1-C3d3 by Lipofectamine 2000. Indirect immunofluorescent assay and Western blot assay showed that VP1-C3d3 fusion gene was successfully expressed in HeLa cells. The fusion protein with the expected size 133 kD could be secreted outside the cells. This study laid a good foundation to further research on the novel vaccine against foot-and-mouth disease virus by using goat C3d as a molecular adjuvant to enhance the immunogenicity of VP1.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes de Fusão
/
Cabras
/
Células HeLa
/
Transfecção
/
Complemento C3d
/
Clonagem Molecular
/
Vírus da Febre Aftosa
/
Proteínas do Capsídeo
/
Alergia e Imunologia
Limite:
Animais
/
Feminino
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2008
Tipo de documento:
Artigo
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