Objective: To construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.
Methods: In line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.
Results: A mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.
Conclusion: The improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.
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