[Expression and purification of GST-Rta fusion protein from EB virus and preparation of the polyclonal antibody against GST-Rta].
OBJECTIVE
To obtain the recombinant fusion proteins GST-Rta185 and GST-Rta150 from EB virus and prepare two Rta protein specific polyclonal antibodies, respectively.


METHODS
Plasmids pGEX-R1501, pGEX-R1851 and pGEX-5X-3 were separately transformed into Escherichia coli BL21 (DE3). Expressions of the recombinant proteins R150-GST, R185-GST and free GST were induced by 0.1 mmol/L IPTG in LB medium. The expressed proteins were purified from lysates with Glutathione Sepharose 4B. Purified proteins were mixed with Freund's adjuvant and then were used to immunize rabbits.


RESULTS
High levels of expression of target proteins were detected in the lysates and the purified proteins were obtained by affinity chromatography with Glutathione Sepharose 4B. Western blot and ELISA analysis suggested that the polyclonal antibodies against GST-R185 and GST-R150 were specific.


CONCLUSIONS
The antiserums have good specificity. They are important for the research on Rta fusion proteins from EB virus and for the diagnosis or treatment of EB virus associated diseases.
Xu
Yong-Chun
YC
Department of Forensic Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Liang
Wei-Bo
WB
Gou
Hang
H
Chen
Wen-Jie
WJ
Zhu
Yin-Hua
YH
Jia
Jing
J
Zhang
Lin
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D020031:Epstein-Barr Virus Infections / Q000821:virology; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D000911:Antibodies, Monoclonal; C068382:BRLF1 protein, Human herpesvirus 4; D017874:Immediate-Early Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D005982:Glutathione Transferase