Cloning and expression of HSV-I, II type-common antigen gD in Escherichia coli / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 176-178, 2002.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-278984
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>BACKGROUND</b>To clone the type common antigen gD of human herpes simplex virus I, II (HSV-I, II), the authors constructed recombinant expression vector Pmal-c2/gD and induced to express the fusion protein MBP-gD.</p><p><b>METHODS</b>The authors extracted HSV DNA,amplified gD gene by PCR assay and directly cloned it into prokaryotic expression vector pMAL-c2, then transformed it into E.coli DH5alpha. After proved to be correct by PCR, double enzyme digestion and sequencing, the fusion protein is induced to express by IPTG and detected by both Western blot and ELISA.</p><p><b>RESULTS</b>The constructed expression vector pMAL-c2/gD can be expressed with high efficiency. The product expressed was about 35.5% of the total bacterium proteins by SDS?PAGE analysis and was found nearly 39% as soluble protein,61% as inclusion in cytoplasm.</p><p><b>CONCLUSIONS</b>The authors constructed recombinant expression vector pMAL-c2/gD, the Western blotting result showed that the recombinant protein could be identified with gD specific monoclonal antibody DL6. Therefore the protein was of natural antigenic structure of gD.</p>
Texto completo:
Disponível
Contexto em Saúde:
Doenças Negligenciadas
Problema de saúde:
Doenças Negligenciadas
/
Zoonoses
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes de Fusão
/
Proteínas do Envelope Viral
/
Clonagem Molecular
/
Escherichia coli
/
Genética
/
Metabolismo
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2002
Tipo de documento:
Artigo