猪圆环病毒2型ORF2基因的定点突变及其真核表达载体的构建

›› 2011, Vol. 38 ›› Issue (2): 99-102.

• 生物技术 • Previous Articles Next Articles

Site-directed Mutagenesis of PCV2 ORF2 Gene Based on Overlap Extension PCR and Construction of Eukaryotic Expression Vector

ZHANG Yan-ping,SU Dan-ping,ZHANG Yan-ping,SU Dan-ping,HE Dong-sheng

（Guangdong Key Laboratory of Zoonosis,College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China）

Received:1900-01-01 Revised:1900-01-01 Online:2011-02-20 Published:2011-02-20
Contact: HE Dong-sheng

Abstract

Abstract: Based on the ORF2 sequences of PCV2 available in GenBank,two pairs of primers were designed and site-directed mutagenesis method based on overlap extension PCR was used to introduce mutations in the ORF2 fragment.DNA sequencing showed that TCTAGAT of 372—378 bp sites had been changed into TATAACT from mutagenesis, the endonuclease sites XbaⅠwas destroyed, site-directed mutagenesis was successfully implemented.Eukaryotic expression vector（pAD5-Blue） was constructed and expressed biological activity protein of Cap.It laid the foundation for further study and played a great role for the PCV2 subunit vaccine research.

Key words: PCV2; ORF2 gene; overlap extension PCR; pAD5-Blue; protein of Cap

CLC Number:

ZHANG Yan-ping;SU Dan-ping;ZHANG Yan-ping;SU Dan-ping;HE Dong-sheng. Site-directed Mutagenesis of PCV2 ORF2 Gene Based on Overlap Extension PCR and Construction of Eukaryotic Expression Vector[J]. , 2011, 38(2): 99-102.

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Site-directed Mutagenesis of PCV2 ORF2 Gene Based on Overlap Extension PCR and Construction of Eukaryotic Expression Vector

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