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| Construction of the High Efficiency and Low Background T Vector |
| HUANG Wen-jun, WANG Ying |
| Key Laboratory of Plant Germplasm Enhancement and Speciality Agriculture,Wuhan Botanical Garden, Chinese Academy of Sciences,Wuhan 430074,China |
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Abstract 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.
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Received: 20 August 2010
Published: 25 December 2010
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