Construction of T-Vector with High Efficiency and Low Background based on the ccdB Killer Gene

Key words: 
• high efficiency /
• low background /
• ccdB gene /
• pMD19-T Simple Vector /
• T-vector

Abstract: A simple and effective method for construction of the T-vector with high efficiency and low background was developed under the universal conditions in laboratory for improving the efficiency of PCR products cloning.The ccdB gene fragment containing restriction enzyme cleavage site was inserted as a medium DNA to the vector backbone p MD19-T Simple. The recombinant plasmid was confirmed by PCR identification,digestion pattern and sequencing,and the recombinant plasmid can then produce a liner T-vector by Xcm I restriction enzyme digestion. This T-vector reduced the background interference of vector self-ligation,and improved the efficiency of T-A cloning with high positive cloning rate. The vector had not only high efficiency and low background but also many advantages of p MD19-T Simple vector,and its preparation was quite simple and convenient for molecular cloning.