[Expression,  purification and activity identification of ZtaN-p23 fusion protein in Escherichia coli of Epstein-Barr virus].
OBJECTIVE
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.


CONCLUSIONS
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.
Shenzhen People's Hospital, Jinan University, Shenzhen, China. zhangyi870907@yahoo.cn
Yi
Y
Zhang
Shu-yan
SY
Zhou
Ye-hong
YH
Sun
Hui-ying
HY
Tu
Xiao-lin
XL
Qin
Rui
R
Chen
Ti-yuan
TY
Li
D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D012995:Solubility; D015534:Trans-Activators / Q000235:genetics* / Q000276:immunology*
C063676:BZLF1 protein, Herpesvirus 4, Human; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators