[Prokaryotic expression, purification and identification of the recombinant gp90 protein of Reticuloendotheliosis virus].
OBJECTIVE
To obtain the recombinant gp90 protein of Reticuloendotheliosis virus (REV) and the anti-gp90 serum with high titer.


METHODS
Using the plasmid pMD18T-env as template, we amplified the gp90 gene and then cloned it into pET-28a(+). The recombinant plasmid pET28a-gp90 was transformed into Escherichia coli BL21 (DE3), which was induced with isopropylthio-beta-D-galactoside(IPTG). After identification by SDS-PAGE and Western blotting, the purified gp90 protein was injected into Balb/c mice to prepare anti-gp90 serum. The specificity and titer of the antiserum were evaluated by IFA and the enzyme-linked immunosorbant assay (ELISA).


RESULTS
SDS-PAGE and Western blotting showed that the gp90 protein was expressed successfully in the form of inclusion body in the recombinant E coli. ELISA showed the mouse anti-gp90 serum had a titer of 1:12800. Successful expression of recombinant gp90 protein and preparation of its antiserum laid the foundation for the development of diagnostic reagent of REV.
Gao
Li
L
Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy Agricultural Sciences, Harbin 150001, China. gaoli0820@163.com
Qi
Xiaole
X
Gao
Honglei
H
Gao
Yulong
Y
Qin
Liting
L
Sun
Fenfen
F
Zhang
Yun
Y
Wang
Xiaomei
X
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D001356:Reticuloendotheliosis virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins