Construction of the High Efficiency and Low Background T Vector
Wenjun Huang, Ying Wāng · Zhongguo shengwu gongcheng zazhi · 2010
A universal method of the high efficiency and low background T vector construction was developed.The gateway cassette fragment containing the ccdB killer gene,used as medium DNA,was ligated to the vector backbone pGEM-T easy,and then the recombinant plasmid was digested by XcmI restriction enzyme to produce the T vector.The recombinant plasmid was confirmed by digestion pattern,PCR identification and sequencing,and the T vector was proved to be high efficiency and low background.This T vector had not only many advantages of pGEM-T easy vector,but also had very high efficiency and low background for molecular cloning.What's more,several restriction enzymes introduced and gateway technology with LR recombination reaction provided convenience for insert fragment subcloning.