中山大学学报自然科学版 ›› 2017, Vol. 56 ›› Issue (2): 1-4.

• 研究论文 •    下一篇

靶向家蚕丝素重链基因的锌指蛋白结构域的人工构建

刘婷婷,梁梓强,梁安文,郭技星,李广宏,王方海   

  1. 中山大学有害生物控制与资源利用国家重点实验室//昆虫学研究所,广东 广州510275
  • 收稿日期:2016-08-10 出版日期:2017-03-25 发布日期:2017-03-25
  • 通讯作者: 王方海(1965年生),男;研究方向:生物化学与分子生物学;E-mail:lsswfh@mail.sysu.edu.cn

Artificial construction of zinc finger protein domain targeting Bombyx mori fibroin heavy chain gene

LIU Tingting,LIANG Ziqiang, LIANG Anwen, GUO Jixing, LI Guanghong, WANG Fanghai   

  1. State Key Laboratory for Biocontrol and Institute of Entomology, Sun Yatsen University, Guangzhou 510275, China
  • Received:2016-08-10 Online:2017-03-25 Published:2017-03-25

摘要:

将多个可特异识别并结合DNA的锌指单体串联在一起即可构成锌指蛋白结构域,若将其与不同的功能域因子融合,则会构建出有很多用途的人工蛋白分子,从而实现对基因组的各种定点修饰或调控。以家蚕丝素重链基因为研究对象,首先利用Zinc Finger Tools软件筛选到了一个合适的靶标位点,再采用模块组装技术,通过酶切、片段回收、连接、转化等步骤成功将多个锌指单体串联在一起,构建出一对靶向家蚕丝素重链基因特定位点的由4个锌指单体组成的锌指蛋白结构域,这为将来定点修饰或改造家蚕丝素基因,进一步提高蚕丝质量和产量,以及大幅度提升家蚕生物反应器对外源蛋白的表达能力打下了良好的基础。

关键词: 家蚕, 丝素重链基因, 锌指蛋白

Abstract:

Many zinc finger units, which can specifically recognize and bind to DNA, are strung together to form zinc finger protein domain. There are many uses to artificially construct the protein molecules, such as various site modification or regulation of the genome, by its integration with the different functional domains of active factors. This study  focused on fibroin heavy chain gene of silkworm. Firstly, a suitable target site was found using Zinc Finger Tools software, and then we used module assembled technology to construct a pair of four zinc finger domain targeting the specific site of fibroin heavy chain gene, through restriction enzyme digesting, fragment recycling, linking, transforming, and other steps. The results would provide a good foundation for the fixedpoint modification or reconstruction of fibroin genes to improve silk quality and production, or to greatly enhance the expression of exogenous protein in the bioreactor of Bombyx mori.

Key words: silkworm, fibroin heavy chain gene, zinc finger protein

中图分类号: