鸡生长激素在毕赤酵母中的表达、纯化和活性鉴定

doi:10.16431/j.cnki.1671-7236.2016.11.024

Abstract

Abstract:

The growth hormone（GH）regulated diverse functions of the target cells through binding its receptor and played important roles in the process of metabolism of the body's growth and development.In order to obtain high purity chicken growth hormone（cGH）with biological activity,the gene segment of the mature cGH with six histidine at the C terminus was cloned into the expression vector pPIC9K.The recombinant plasmid（pPIC9K-cGH）was transformed to Pichia pastoris（GS115）by electroporation to construct the recombinant Pichia pastoris（GS115-pPIC9K-cGH).The GS115-pPIC9K-cGH secreted the recombinant protein（cGH）whose molecular weight was about 25 ku induced by 1% methanol induction for 96 h.The purity of cGH attained at least 95% through using Ni affinity chromatography and polyacrylamide glucan gel chromatography to purify the recombinant protein.The results of Real-time quantitative-PCR showed that the purified cGH recombinant proteins could up-regulate its target gene insulin-like growth factor Ⅰ（IGF-Ⅰ）mRNA in chicken liver cancer cell（LMH).It certified that the recombinant cGH protein had biological activity.This study laid a foundation for research of the structure,function and application of cGH.

Key words: chicken growth hormone; Pichia pastoris; expression; purification; biological activity

CLC Number:

S831.2

YU Jian-feng, HE Sai, JIN Ze-kai, ZHOU Huan-qing, DONG Xiao-min, GONG Dao-qing, GU Zhi-liang. Expression,Purification and Biological Activity Identification of Chicken Growth Hormone in Pichia pastoris[J]. , 2016, 43(11): 2970-2975.

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Expression,Purification and Biological Activity Identification of Chicken Growth Hormone in Pichia pastoris

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