A Novel PCR Site-Directed Mutagenesis to Modify Structure of Erabutoxin B

Fang Hui Wu; Yan Jun Li; Ting Song Yang; Tian Yi Ying

doi:10.4028/www.scientific.net/AMR.183-185.605

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HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 183-185A Novel PCR Site-Directed Mutagenesis to Modify...

A Novel PCR Site-Directed Mutagenesis to Modify Structure of Erabutoxin B

Abstract:

Objective: A convenient PCR was used for site-directed mutagenesis to modify structure of proteins or peptides, For function-structure studies of proteins or peptides. Method: the one-step PCR site-directed mutagenesis strategy can introduce mutation of gene through 5^/-end of primer, which was applied on peptide Erabutoxin B (EB) to produce three mutants of EB, the activity of mutants was detected by LD50 value of mice. Result: S8Y, R33D, K47R three mutants of EB were obtained by one-step PCR, LD₅₀ of mutants indicated that the activity of mutants decreased in different degree, the activity of R33D was nearly deprived. Conclusion: one-step PCR site-directed mutagenesis was convenien and efficient, it can be applied on restructuring the primary structure of proteins or peptides.

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Periodical:

Advanced Materials Research (Volumes 183-185)

Pages:

605-610

DOI:

https://doi.org/10.4028/www.scientific.net/AMR.183-185.605

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Online since:

January 2011

Authors:

Fang Hui Wu, Yan Jun Li, Ting Song Yang, Tian Yi Ying

Keywords:

Erabutoxin B, LD₅₀, One-Step PCR, Site-Directed Mutagenesis

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