大肠杆菌NrfA蛋白表达_纯化及多克隆抗体的制备_英文_.pdfVIP

Agricultural Science Technology, 2012, 13(4): 723-726 Copyright 訫 2012, Information Institute of HAAS. All rights reserved Agricultural Biotechnology Expression and Purification of E. coli NrfA Protein and Preparation of Polyclonal Antibody against NrfA ， 1 2 1 * 3 Tingting HE , Gangming GONG , Ran GAO 1. School of Perfume and Aroma Technology, Shanghai Institute of Technology, Shanghai 200235, China; 2. College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China; 3. Institute of Biochemistry, Zhejiang Sci-Tech University, Hangzhou 310018, China Materials and Methods Abstract [Objective] This study aimed to clone the E. coli NrfA gene and construct the pET-28a (+)-NrfA prokaryotic expression vector for preparation of polyclonal anti- Materials body against E. coli NrfA. [Method] E. coli NrfA gene was cloned from the E. coli Experimental animals New Zealand genome DNA by PCR and inserted into the vector pET-28a(+) to construct prokary- male rabbit about 3 kg in weight was otic expression vector pET-28a (+)-NrfA. E. coli NrfA protein was expressed by IPTG purchased from Tianjin Aoyi Animal induction and purified. Polyclonal antibody against NrfA protein was prepared by im- munizing rabbit with routine method. The specificity and titer of polyclonal antibody Breeding Center. was confirmed by ELISA and Western Blotting. [Result] The constructed prokaryotic Reagents Peptone, beef extract and expression ve