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| A Cloning Strategy Suitable for DNA Modification by Fragment Scanning |
| GAN Chun-yang, LIU Ya, LUO Ying-ying, ZHANG Wen-lu, HUANG Ai-long, CAI Xue-fei, HU Jie-li |
| Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China |
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Abstract DNA sequence modification is a basic technology in genetic engineering. One aspect of DNA modification is insertion or substitution with large heterogeneous fragments. Sometimes fragments need to be inserted or substituted to a sequence with a scanning model, to obtain an optimal sequence meeting specific purpose. Traditional cloning method using restriction digestion and ligation reactions might become laborious and time-consuming, or even unpractical in these cases. A strategy combining Golden gate method, mixing cloning, and rapid identification of clone (named Gmix) was reported. By using Gmix, the Gaussia gene was inserted or substituted to specific location in a HBV replication plasmid with 4 different scanning models. With high successful rate, easy handling and low cost, Gmix is suitable for many other DNA modification works resembling that described here.
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Received: 29 January 2016
Published: 25 August 2016
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