[Expression, purification and activity identification of ZtaN-p23 fusion protein in Escherichia coli of Epstein-Barr virus].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 28(2): 120-3, 2012 Feb.
Article
em Zh
| MEDLINE
| ID: mdl-22304765
ABSTRACT
AIM:
To construct the prokaryotic expression plasmid pGEX-4T-1-BZLF1N-BLRF2, and express it in Escherichia coli.METHODS:
The EB virus BZLF1N gene and BLRF2 gene were amplified by RT-PCR respectively. Then, the two genes were linked by splicing overlap extension PCR method and inserted into the vector pGEX-4T-1, and the recombinant plasmid pGEX-4T-1-BZLF1N-BLRF2 was transformed into E.coli BL21 (DE3) strain. The expression protein ZtaN-p23 was analysed by SDS-PAGE and immunoreactivity was proved by Western blotting.RESULTS:
Restriction enzyme digestion and DNA sequencing showed recombinant plasmid constructed successfully. The expression product ZtaN-p23 with the molecular weight 46000 was located in the cytoplasm and insoluble. The ZtaN-p23 up to 95% purity was obtained after purified using affinity chromatography. Western blotting showed fusion protein possessed a well bioactivity and specificity.CONCLUSION:
The fusion gene BZLF1N-BLRF2 is successfully constructed and effectively expressed in E.coli, which lay the foundation for further research on its biological properties and functions.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes de Fusão
/
Expressão Gênica
/
Transativadores
/
Herpesvirus Humano 4
/
Escherichia coli
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
Zh
Ano de publicação:
2012
Tipo de documento:
Article