Human CD96 gene cloning, expression and identification / 南方医科大学学报
Journal of Southern Medical University
; (12): 1232-1235, 2011.
Article
em Zh
| WPRIM
| ID: wpr-235155
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.</p><p><b>METHODS</b>hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.</p><p><b>RESULTS</b>hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.</p><p><b>CONCLUSION</b>The CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.</p>
Texto completo:
1
Base de dados:
WPRIM
Assunto principal:
Células-Tronco Neoplásicas
/
Proteínas Recombinantes
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Leucemia Mieloide Aguda
/
Antígenos CD
/
Imunização
/
Clonagem Molecular
/
Alergia e Imunologia
/
Escherichia coli
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Journal of Southern Medical University
Ano de publicação:
2011
Tipo de documento:
Article