Prokaryotic expression and purification of human immunodeficiency virus p24 antigen / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 28-31, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-333058
ABSTRACT
<p><b>OBJECTIVE</b>To express and purify the HIV p24 gene in E. coli cells, and identify p24 antigen activity.</p><p><b>METHODS</b>The full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET-p24 recombined vector. After transforming into E. coil, the purified p24 protein was prepared by metal-ligand affinity chromatography (IMAC). The accuracy of inserted gene and activity, specificity of HIV p24 proteins were detected by two enzymes digestion technology, SDS-PAGE, Western Blot (WB) and ELISA.</p><p><b>RESULTS</b>The length of the HIV p24 gene fragment was 690 bp after digesting the recombinant plasmid T-p24 and pRSET-p24 with BamH I and Hind III. The expressed proteins had a single expected band of about 24 x 10(3) in SDS-PAGE. The specificity and activity of p24 protein were tested by WB and ELISA.</p><p><b>CONCLUSION</b>The HIV p24 sequence from HIV-1 gene plasmid hasbeen expressed in E. coil. This protein possessed good specificity and activity.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes
/
Ensaio de Imunoadsorção Enzimática
/
Expressão Gênica
/
Western Blotting
/
Reação em Cadeia da Polimerase
/
Cromatografia de Afinidade
/
Proteína do Núcleo p24 do HIV
/
Clonagem Molecular
/
Eletroforese em Gel de Poliacrilamida
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2005
Tipo de documento:
Artigo
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