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

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 to 378 bp sites had been changed into TATAACT from mutagenesis,the endonuclease sites XbaⅠwas 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.