QKI protein in prokaryotic expression purification and polyclonal antibody preparation and characterization of.doc

 PAGE \* MERGEFORMAT 14 QKI protein in prokaryotic expression purification and polyclonal antibody preparation and characterization of Authors: HUANG, Yu-fang, WANG Xiao-Gong, Bai Li Yuan, Li Hua, Lutz Fan [Keywords:] QKI [Abstract] Objective: To obtain the original expression of the RNA-binding protein QKI7 protein and preparation of its polyclonal rabbit anti-QKI antibodies. Methods: The cDNA coding region were successfully inserted into QKI7 the PET32b () prokaryotic expression vector was transformed by heat shock BL21 (DE3) Competent bacteria to IPTG induced 6 * HisQKI7 fusion protein expression, respectively, by metal-chelating column, the reverse column and strong anion-exchange chromatography column for purification. By SDSPAGE and Western blot identified, application of purified fusion protein, respectively Freund’s adjuvant, polyacrylamide gel, NC membrane as an adjuvant, prepared by immunization of New Zealand white rabbits polyclonal antibodies, and to Western blot testing. Results: The expression and purification of QKI7 protein more than 95% purity, the application of purified fusion protein as an adjuvant to Freund’s adjuvant immunization of New Zealand white rabbits, was a high titer of anti-QKI-specific polyclonal antibody. Conclusion: The successful expression and purification of QKI7 protein and preparation of high titer and high specificity of anti-QKI polyclonal antibody. With Freund’s adjuvant as an immune adjuvant for immunization is better than the polyacrylamide gel and the NC membrane. [Keywords:] QKI7; prokaryotic expression; protein purification; polyclonal antibody QKI belongs to a kind of STAR family RNA-binding protein, highly conserved during evolution, in the nervous system myelin formation, embryonic development, growth and other aspects of the cardiovascular system plays an important role. qki genes in different parts of mutant mice die before birth can be expressed in cardiovascular muscle system developmental disorders,