Reliable Scarless Fusion of Genes through Overlap Extension PCR to Develop Chimeric Genes
Reliable Scarless Fusion of Genes through Overlap Extension PCR to Develop Chimeric Genes
Iram Gull*, Muhammad Shahbaz Aslam, Imran Tipu, Roohi Mushtaq and Muhammad Amin Athar
ABSTRACT
Overlap extension PCR is used to make versatile chimeric gene products. However, the results of this technique are not always reliable due to inefficient overlapping of the sequences especially those derived from authenticated native sequences. Certain forbidden sequences do appear at junction of spliced genes whenever efficiency of gene fusion is improved by inserting GC rich sequences. In this study, reliable scar-less gene fusion with the insertion of sequence of choice at junction sites has been described by optimization of various parameters of overlap extension PCR. The fusion of IFNα2b gene to the gene of human latency associated protein with insertion of HCV NS3 protease cleavage site at splicing junction is reported.
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August 2019
Vol. 51, Iss. 4, Pages 1203-1598
