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凡纳滨对虾抗菌肽crunstinA在毕赤酵母菌中的表达

马春霞 彭金霞 何苹萍 雷爱莹 马宁 王瑞 黎铭

南方农业学报2017,Vol.48Issue(7):1310-1316,7.
南方农业学报2017,Vol.48Issue(7):1310-1316,7.DOI:10.3969/j.issn.2095-1191.2017.07.30

凡纳滨对虾抗菌肽crunstinA在毕赤酵母菌中的表达

Expression of Litopenaeus vannamei antimicrobial peptide crunstinA in Pichia pastoris

马春霞 1彭金霞 2何苹萍 2雷爱莹 2马宁 2王瑞 2黎铭2

作者信息

  • 1. 广西兽医研究所, 南宁 530001
  • 2. 广西水产科学研究院/广西遗传育种与健康养殖重点实验室, 南宁 530021
  • 折叠

摘要

Abstract

[Objective]Expression of antimicrobial peptides crunstinA(LvcrustinA) isolated from Litopenaeus van-namei in Pichia pastoris was studied in order to lay a foundation for developing new aquaculture medicines based on re-combinant protein crunstinA. [Method]LvcrustinA gene was synthesized by PCR-based accurate synthesis(PAS) method. LvcrustinA gene was cloned into eukaryotic expression vector pYE-GAPα. The recombinant plasmid obtained transformed into competent bacteria TOP10. Recombinanted plasmid pYE-GAPα-Lvcrustin-A extracted by plasmid extraction kit was transformed into P. pastoris GS115. Expression of fusion protein LvcrustinA was analyzed by SDS-PAGE electrophoresis detection and Western blotting identification. Resistance effects of fusion protein LvcrustinA to Vibrio parahaemolyticus were tested by protection experiment. [Result]The established recombinant plasmid pYE-GAPa-LvcrustinA was trans-formed into P. pastoris strain GS115, the obtained recombinant strain could be cultured in shaking table(220 r/min) for 24 h under 30 ℃ to produce fusion protein LvcrustinA. As the culture time extended, the expression increaseed. Western blotting identification indicated that purified fusion protein LvcrustinA could be identified by His antibody. Protection ex-periment results showed that fusion protein LvcrustinA was resistant to V. parahaemolyticus. [Conclusion]Expression plasmid pYE-GAPa and P. pastoris strain GS115 can recombine into LvcrustinA. LvcrustinA can help L. vannamei resist infection of V. parahaemolyticus.

关键词

凡纳滨对虾/抗菌肽/毕赤酵母菌/真核表达

Key words

Litopenaeus vannamei/antimicrobial peptides/Pichia pastoris/eukaryotic expression

分类

农业科技

引用本文复制引用

马春霞,彭金霞,何苹萍,雷爱莹,马宁,王瑞,黎铭..凡纳滨对虾抗菌肽crunstinA在毕赤酵母菌中的表达[J].南方农业学报,2017,48(7):1310-1316,7.

基金项目

广西自然科学基金项目(2015GXNSFAA139067) (2015GXNSFAA139067)

广西水产畜牧科技推广应用项目(桂渔牧科201528012) (桂渔牧科201528012)

南方农业学报

OA北大核心CSCDCSTPCD

2095-1191

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