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- W2377884823 abstract "Objective: A convenient and rapid site-directed mutagenesis method was established for study regulation of gene expression as well as relationship between protein structure and function.Method:The construction of bsh(encoding bile salt hydrolase in Listeria monocytogenes) promoter mutants was used as a sample in this study.A pair of completely complementary primers with mutation sites in the middle was used to amplify the total recombinant plasmid DNA sequences.After digestion of residual methylated template DNA in the PCR products by DpnⅠ,PCR products without purification were directly transformed into E.coli to contain mutations of the recombinant plasmid.Result: Three bsh promoter mutants were successfully constructed by the one-step site-directed mutagenesis technology.Conclusion: The method used in this study is robust and fast.As long as primer design,high-fidelity DNA polymerase,template DNA concentration,and PCR amplification procedure are optimized,the successful rate of mutation could reach 100%." @default.
- W2377884823 created "2016-06-24" @default.
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- W2377884823 date "2009-01-01" @default.
- W2377884823 modified "2023-09-23" @default.
- W2377884823 title "Construction of bsh Promoter Mutants by Rapid One-step Site-directed Mutagenesis" @default.
- W2377884823 hasPublicationYear "2009" @default.
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