Biotechnology Bulletin ›› 2016, Vol. 32 ›› Issue (3): 209-214.doi: 10.13560/j.cnki.biotech.bull.1985.2016.03.033

• Research report • Previous Articles     Next Articles

Expression of Protein Us11 and Preparation of Polyclonal Antibodies

TONG Hai-yan1, LI Guo-yi1, 2, TANG Ying1   

  1. 1. The First Affiliated Hospital of Harbin Medical University, Harbin 150001;
    2. Oriental Hospital Affiliated to Tongji University, Shanghai 200085
  • Received:2015-05-28 Online:2016-03-24 Published:2016-03-25

Abstract: The previous studies have discovered that protein Us11 of HSV-1(herpes simplex virus 1)played an important role in antagonistic host innate immune responses after HSV-1 infection. By synthesizing the gene Us11 of HSV-1, the polyclonal antibodies for Us11 were cloned and prepared, which provided the solid experimental basis for studying the functions of Us11 and the interactions between Us11 and host protein. The artificially-synthesized gene Us11 was as template and amplified, and then the recycled gene fragments of Us11 were cloned into the prokaryotic and eukaryotic expression vectors. Further, the induced expressed Us11 protein was purified, and polyclonal antibodies for rabbit were prepared. The recombinant plasmid was transfected into 293T cells;the expressed protein was detected by indirect immunofluorescence(IFA)and Western blot(WB). Plasmid pCold-Us11 and pFLAG-Us11 were successfully constructed, soluble Us11 protein expression was achieved, and the polyclonal antibodies targeting for Us11 was acquired by immunizing animal with purified Us11 protein. The results from IFA and WB revealed that prepared antiserum may specially detect the Us11 protein expressed in transfected 283T cells by pFLAG-Us11.

Key words: Us11, prokaryotic expression, eukaryotic expression, polyclonal antibody