摘要
为了适用于NimbleCloning这种新型的DNA分子克隆技术,本研究以pCambia1304载体作为改造对象,采用突变碱基的方式去除载体中的5个SfiI酶切位点,并且对改造后的载体在农杆菌中的稳定性以及在植物中的表达进行分析,结果表明5个SfiI位点突变对载体的稳定性和表达没有影响。本研究为NimbleCloning系统提供了配套载体,也为其他pCambia系列载体改造成NimbleCloning系统载体提供了模板和参考。
In order to construct compatible vectors for Nimble Cloning, a new technology of DNA molecular cloning, the plant vector pCambia1304 was used as a reconstruction target in this study. Five Sfi I restriction sites in the pCambia1304 were removed by base substitution. The stability of the reconstructed vector in Agrobacterium and the expression in plants were analyzed. The results showed that the mutations of 5 Sfi I sites had no negative effect on the stability and expression of the vector. This study provides a compatible vector for the Nimble Cloning system and a template and references for reconstructing other vectors of pCambia series into Nimble Cloning system vectors.
作者
曾妍静
沈文涛
庹德财
黎小瑛
言普
周鹏
ZENG Yan-jing;SHEN Wen-tao;TUO De-cai;LI Xiao-ying;YAN Pu;ZHOU Peng(College of Agriculture,Hainan University,Haikou 570228,Hainan,China;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou 571101,Hainan,China)
出处
《生命科学研究》
CAS
CSCD
2019年第4期276-280,共5页
Life Science Research
基金
中国热带农业科学院基本科研业务费专项(1630052018004,1630052019004)
