Construction of Eukaryotic Expression Vector of Mustela Vision Growth Hormone and Expression in Yeast Pichia Pastoris

Long Yingna1,2, Ma Fenglong2 , Qiao Yanliang2, Liu Huanqi1, Wang Mingzhi1 (1Department of Animal Science and Veterinary Medicine of Qingdao Agricultural University, Qingdao 266109; 2Sinder Medicine Technic Share Limited Company Shandong, Qingdao 266061)

Mustela vision growth hormone; Pichia Pastoris GS115; pPIC9K vector; SDS-PAGE; Western blot

[Objective]To construct eukaryotic expression vector of Mustela vision growth hormone(mGH), and express in Pichia Pastoris strain GS115, make foundation to gain amounts of Mustela vision growth hormone. [Method]Mustela vision growth hormone, which was chemically synthesized, was inserted into secretion vector pPIC9K. The recombined pPIC9K–mGH was digested by Sac I and transferred into Pichia Pastoris strain GS115 with LiC1. After screening with G418, the positive yeast clone was induced to express with methanol, and the expression product was analyzed with SDS-PAGE and Western blot. [Results]A positive protein band with molecular weight about 31 kD was found in the expression product, which is accordant with interest protein, and this band could be specifically recognized by anti-mGH polyclonal antibody. [Conclusion]The gene of mGH was cloned into the expression vector correctly and was expressed successfully in P. pastoris expression system.