Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Cloning, expression and characterization of gE protein of Duck plague virus | Chang et al. Virology Journal 2010 7 120 http content 7 1 120 VIROLOGY JOURNAL RESEARCH Open Access Cloning expression and characterization of gE protein of Duck plague virus Hua Chang11 Anchun Cheng 21 1 2 3 Mingshu Wang21 2 Dekang Zhu21 2 Renyong Jia22 Fei Liu2 Zhengli Chen2 Qihui Luo2 Xiaoyue Chen1 2 3 and Yi Zhou2 Abstract Background The gE protein of duck plague virus is the important membrane glycoprotein its protein characterization has not been reported. In this study we expressed and presented the characterization of the DPV gE product. Results According to the sequence of the gE gene a pair of primers were designed and the DNA product with 1490bp in size was amplified by using the polymerase chain reaction PCR . The PCR product was cloned into pMD18-T vector and subcloned into pET32a generating the recombinant plasmid pET32a DPV-gE. SDS-PAGE analysis showed that the fusion pET32a DPV-gE protein was highly expressed after induction by mM IPTG at 30 C for h in Rosseta host cells. Over expressed 6xHis-gE fusion protein was purified by nickel affinity chromatography and used to immunize the rabbits for the preparation of polyclonal antibody. The result of the intracellular localization revealed that the gE protein was appeared to be in the cytoplasm region. The real time PCR RT-PCR analysis and Western blotting revealed that the gE gene was produced most abundantly during the late phase of replication in DPV-infected cells. Conclusions In this work the DPV gE protein was successfully expressed in a prokaryotic expression system and we presented the basic properties of the DPV gE product for the first time. These properties of the gE protein provided a prerequisite for further functional analysis of this gene. Background Duck plague DP which is caused by DPV is an acute febrile contagious and septic disease of waterfowl ducks geese and swans 1 . DPV has been classified as belonging to the Alphaherpesvirinae subfamily of the family .
