›› 2005, Vol. 48 ›› Issue (2): 179-183.doi:

• 研究论文 • 上一篇    下一篇


刘忠渊,张富春*,王 芸,吕国栋   

  1. 新疆大学生命科学与技术学院分子生物学重点实验室,新疆生物资源基因工程重点实验室
  • 出版日期:2005-07-10 发布日期:2005-11-20
  • 通讯作者: 张富春

Expression of the antifreeze protein gene of a pyrochroid beetle Dendroides canadensis in prokaryote and detection of the protein biological activity

LIU Zhong-Yuan, ZHANG Fu-Chun*, WANG Yun, LU Guo-Dong   

  1. Key Laboratory of Molecular Biology, College of Life Science and Technology
  • Online:2005-07-10 Published:2005-11-20
  • Contact: ZHANG Fu-Chun

摘要: 根据GenBank中序列人工合成赤翅甲Dendroides canadensis的抗冻蛋白基因(afp),将其克隆到载体pGEX-4T-1上,构建融合表达的重组质粒,转化大肠杆菌 BL21并进行原核表达。通过优化表达的诱导条件和SDS-PAGE检测,证明人工合成的赤翅甲抗冻蛋白基因能够特异性地表达,并以可溶性融合蛋白形式存在,相对分子质量约为40 kD。抗冻蛋白的生物活性检测表明,赤翅甲的抗冻融合蛋白能够提高细菌的耐寒能力。

关键词: 赤翅甲, 抗冻蛋白, 原核表达, 抗冻活性

Abstract: In order to express a pyrochroid beetle Dendroides canadensis antifreeze protein in E. coli BL21 and to investigate the properties of recombinant antifreeze protein, the D. canadensis afpgene was synthesized,  cloned into the pGEX-4T-1, and transformed into E. coli BL21 The optimal inductive condition was selected by  different experimental approaches. The results of SDS-PAGE analysis indicated that the antifreeze protein was expressed successfully in E.coli BL21 The expressed antifreeze protein, existing in the soluble and fusion form, with the molecular weight of about 40 kD, could increase low temperature resistance of bacteria as shown by the biological activity detection.

Key words: Dendroides canadensis, antifreeze protein, prokaryotic expression, biological activity