The PCR production of vascular endothelial growth factor (VEGF) was subcloned into a T-vector, and the recombined plasmids were then transformed into competent cell JM109. Eight white clones were selected randomly, and the recombined plasmids were extracted and mixed as template. Three selected primers were adapted and two rounds of PCR were performed to introduce mutations. The production of those were confirmed by enzyme digestion, PCR amplification and sequencing. The method is proved to be a simple,quick and easy way to introduce site-directed mutagenesis, and make latter work easy to do.
BAI Xiang-Yang, Lü An-Guo, WU Wen-Fang, SUN Jing, NIU Rui-Fang. Site-Directed Mutagenesis of Vascular Endothelial Growth Factor (VEGF) by An Improved Overlap PCR[J]. Progress in Biochemistry and Biophysics,2004,31(2):181-184
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