食品科学 ›› 2009, Vol. 30 ›› Issue (5): 126-129.doi: 10.7506/spkx1002-6630-200905029

• 生物工程 • 上一篇    下一篇

鲑鱼降钙素与降钙素基因相关肽融合基因植物表达载体的构建

余 琼,王 超,李荣田,平文祥,孙怡宁,李洪丽   

  1. 黑龙江省高等学校分子生物学重点实验室;黑龙江大学生命科学学院
  • 收稿日期:2008-03-20 修回日期:2008-10-28 出版日期:2009-03-01 发布日期:2010-12-29
  • 通讯作者: 余琼 E-mail:yuqiong00001@163.com
  • 基金资助:

    黑龙江省教育厅2007 年科学技术研究面上项目(11521223) 。

Construction of Plant Expression Vector of Salmon Calcitonin (sCT) and Calcitonin Gene-related Peptide (CGRP) Fusion Gene

YU Qiong,WANG Chao,LI Rong-tian,PING Wen-xiang,SUN Yi-ning,LI Hong-li   

  1. (Key Laboratory of Molecular Biology, College of Heilongjiang Province, College of Life Science, Heilongjiang University, Harbin
    150080, China)
  • Received:2008-03-20 Revised:2008-10-28 Online:2009-03-01 Published:2010-12-29
  • Contact: YU Qiong E-mail:yuqiong00001@163.com

摘要:

根据基因工程操作方法和植物偏爱密码子原则,设计了鲑鱼降钙素与降钙素基因相关肽融合基因,并在融合基因前端加上AMV RNA4 5'-UTR 序列和6 × His 标签,在融合基因后边加上一个甘氨酸。通过Sac I 和Sma I双酶切pUC(AMV / CGRP / msCT)获得融合基因,用T4 DNA 连接酶连到植物表达载体pBI121 上,构建重组质粒pBI AMV / CGRP / msCT,再转化到E.coli DH5α中保存。通过DNA 测序验证表达载体构建成功。

关键词: 鲑鱼降钙素, 降钙素基因相关肽, 基因工程, 融合表达, 高血压, 骨质疏松

Abstract:

The fusion gene of sCT and CGRP was designed according to the principles of genetic engineering method and the bias codon used by plant. An AMV RNA4 5'-UTR sequence and a 6×His tag were added before the gene, and the Gly was added at the end of the fusion gene. The fusion gene was obtained from pUC(AMV / CGRP / msCT) by Sac I and Sma I digestion, then cloned into plant expression vector pBI121 by T4 DNA ligase to construct recombinant plasmid pBI AMV / CGRP / msCT, and transformed into E.coli DH5α to be kept. The gene sequencing indicated that the expression vector pBI AMV / CGRP / msCT has been successfully constructed.

Key words: salmon calcitonin, calcitonin gene-related peptide, genetic engineering, fusion expression, hypertension, osteoporosis

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