生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (3): 119-.doi: 10.3969/j.issn.2095-1736.2023.03.119

• 教学研究 • 上一篇    下一篇

本科生基因工程实验课程基因克隆构建方案

陆盈盈， 樊伟康， 韦有恒   

  1. 扬州大学 生物科学与技术学院， 扬州 225009
  • 出版日期:2023-06-18 发布日期:2023-06-19
  • 通讯作者: 韦有恒，博士，教授，研究方向为生物化学，E-mail:yhwei@yzu.edu.cn
  • 作者简介:陆盈盈，硕士，实验师，研究方向为基因工程，E-mail:006550@yzu.edu.cn
  • 基金资助:
    国家自然科学基金面上项目（31872287）

Gene cloning construction method for undergraduate genetic engineering experimental course

LU Yingying, FAN Weikang, WEI Youheng   

  1. College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China
  • Online:2023-06-18 Published:2023-06-19

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摘要： 将常用于Gateway克隆系统中的对大肠杆菌具有毒性ccdB基因引入载体，提高基因克隆成功率。构建基因克隆过程中，含有ccdB 基因的原始载体在大肠杆菌中不能正常生长，无法形成细胞克隆，避免空载的假阳性克隆产生。通过分组比较实验、启发引导的教学设计，可以让学生充分领会酶切、重组、连接等生物工程相关操作，加深学生对理论课程的理解。该基因克隆方法的改进能够满足实验教学的需要，也适用于科研工作中的基因克隆效率的提高。

关键词: 基因工程, 实验教学, 构建基因克隆, ccdB, 质粒

Abstract: The ccdB gene was introduced, using the Gateway system, to the vector to improve the efficiency of clone construction. The Escherichia coli transfected with vectors containing ccdB could not grow to form clones because the ccdB is toxic to it. Thus, this strategy avoided the false positive clone formation and made the plasmid construct much more convenient. Through comparison of the different strategies of clone construction, this class made students understand more on genetic engineering-related processes such as enzyme digestion, recombination, ligation. This method is suitable not only for the experimental course but also for cloning construction in future research.

Key words: genetic engineering, experimental teaching, cloning construction, ccdB, plasmid

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