2009China Animal Husbandry & Veterinary MedicineRequires access

Application of Two Methods for Site-specific Mutagenesis of Porcine IκBα

Yang Shu-lin

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Abstract

Substitute Ala for the sites of Ser 32 and Ser 36 in porcine IκBα gene,to establish the fundament for mechanism research of the mutant IκBα inhibition on delayed xenograft rejection.QuikChange Site-Directed Mutagenesis Kit was first uesd for the mutagenesis of Ser 32 site,then the kit was subsequently used for the mutagenesis of Ser 36 site,but failed.Finally OE-PCR was used for the mutagenesis of Ser 36 site.These two mutagenesis methods were combined to achieve site-specific mutagenesis at the sites of Ser 32 and Ser 36 in porcine IκBα gene.These two methods each has a unique,and each has their own shortcoming.So,in order to get better result by less work,mutation methods should be chosen flexibly according to the real need.

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What this paper is about

Substitute Ala for the sites of Ser 32 and Ser 36 in porcine IκBα gene,to establish the fundament for mechanism research of the mutant IκBα inhibition on delayed xenograft rejection.QuikChange Site-Directed Mutagenesis Kit was first uesd for the mutagenesis of Ser 32 site,then the kit was subsequently used for the mutagenesis of Ser 36 site,but failed.Finally OE-PCR was used for the mutagenesis of Ser 36 site.These two mutagenesis methods were combined to achieve site-specific mutagenesis at the sites of Ser 32 and Ser 36 in porcine IκBα gene.These two methods each has a unique,and each has their own shortcoming.So,in order to get better result by less work,mutation methods should be chosen flexibly according to the real need.

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Available abstract

Substitute Ala for the sites of Ser 32 and Ser 36 in porcine IκBα gene,to establish the fundament for mechanism research of the mutant IκBα inhibition on delayed xenograft rejection.QuikChange Site-Directed Mutagenesis Kit was first uesd for the mutagenesis of Ser 32 site,then the kit was subsequently used for the mutagenesis of Ser 36 site,but failed.Finally OE-PCR was used for the mutagenesis of Ser 36 site.These two mutagenesis methods were combined to achieve site-specific mutagenesis at the sites of Ser 32 and Ser 36 in porcine IκBα gene.These two methods each has a unique,and each has their own shortcoming.So,in order to get better result by less work,mutation methods should be chosen flexibly according to the real need.

Key concepts: Mutagenesis, Site-directed mutagenesis, Mutant, Gene, Mutation, Directed mutagenesis, Insertional mutagenesis, Biology

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