[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. CONCLUSION: 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.