华北大黑鳃金龟丝氨酸羧肽酶在不同体系中的表达

doi:10.7668/hbnxb.2013.05.008

华北农学报 ›› 2013, Vol. 28 ›› Issue (5): 44-47. doi: 10.7668/hbnxb.2013.05.008

华北大黑鳃金龟丝氨酸羧肽酶在不同体系中的表达

张雅昆¹, 赵丹³, 郭巍^1,2,3, 刘小民⁴, 徐大庆¹, 孙伟明³

1 河北农业大学生命科学学院河北保定 071001;
2 河北农业大学植物保护学院河北省农作物病虫害生物防治工程技术中心河北保定 071001;
3 北京农学院植物科学技术学院北京 102206;
4 河北省农林科学院粮油作物研究所河北石家庄 050035

收稿日期:2013-07-17 出版日期:2013-10-28
通讯作者: 郭巍(1971－),女,黑龙江绥化人,教授,博士生导师,主要从事生物化学与分子生物学教学与研究。;徐大庆(1972－),男,河北平泉人,副教授,硕士生导师,主要从事生物化学与分子生物学教学与研究。
作者简介:张雅昆(1988－),女,河北保定人,硕士,主要从事昆虫生物化学与分子生物学研究。
基金资助:
国家重点基础研究发展规划(“973”计划)项目(2009CB118902);国家自然科学基金项目(30971910);国家现代农业产业技术体系项目

Expression of a Serine Carboxypeptidase HoSCP of Holotrichia oblita( Coleoptera: Scarabaeoidea) in Different Systems

ZHANG Ya-kun¹, ZHAO Dan³, GUO Wei^1,2,3, LIU Xiao-min⁴, XU Da-qing¹, SUN Wei-ming³

1 College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China;
2 Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China;
3 College of Plant Protection, Agricultural University of Hebei, Biological Control Center of Plant Diseases and Plant Pests of Hebei Province, National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071001, China;
4 Institute of Cereal and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050035, China

Received:2013-07-17 Published:2013-10-28

摘要/Abstract

摘要： 丝氨酸羧肽酶(Serine carboxypeptidases,SCPs)是一类在酸性pH环境下参与多肽和蛋白质的加工、修饰与降解等多个重要环节的真核生物蛋白水解酶。根据华北大黑鳃金龟丝氨酸羧肽酶(HoSCP)全长基因序列设计引物,利用PCR方法克隆hoscp基因并分别连接到表达载体pET28b和pPIC9K上,转化大肠杆菌BL21和毕赤酵母GS115,实现了其在原核以及真核体系中的表达。结果显示,HoSCP在原核细胞及酵母细胞中均能稳定表达50 kDa的特异蛋白。HoSCP的成功表达为其在华北大黑鳃金龟体内的活性和生物学功能研究提供了可能。

关键词: 华北大黑鳃金龟, 丝氨酸羧肽酶, HoSCP, 原核表达系统, 毕赤酵母表达系统

Abstract: Serine carboxypeptidases ( SCPs ) are a class of protease enzyme that play essential roles in a wide range of biological processes which are the modification and degradation of peptides and proteins. We report,in this study,the hoscp gene of Holotrichia oblita was amplified by PCR method and cloned into the expression vector pET 28b and pPIC9K,respectively. The following recombinant vector was transformed into Escherichia coli BL21 and Pichia pastoris GS115,respectively. The results showed that HoSCP was stably expressed as a 50 kDa recombinant protein in E.coli and P.pastoris.It is useful for the further study of activity and biological function of HoSCP in H.oblita.

Key words: Holotrichia oblita, SCPs, HoSCP, Prokaryotic expression system, P. pastoris expression system

中图分类号:

S433.5

张雅昆, 赵丹, 郭巍, 刘小民, 徐大庆, 孙伟明. 华北大黑鳃金龟丝氨酸羧肽酶在不同体系中的表达[J]. 华北农学报, 2013, 28(5): 44-47. doi: 10.7668/hbnxb.2013.05.008.

ZHANG Ya-kun, ZHAO Dan, GUO Wei, LIU Xiao-min, XU Da-qing, SUN Wei-ming. Expression of a Serine Carboxypeptidase HoSCP of Holotrichia oblita( Coleoptera: Scarabaeoidea) in Different Systems[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(5): 44-47. doi: 10.7668/hbnxb.2013.05.008.

http://www.hbnxb.net/CN/Y2013/V28/I5/44

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