Site-directed Mutagenesis of pBluescript Ⅱ SK(+) and pCAMBⅠA1300 Vectors Based on PCR Method

doi:10.7668/hbnxb.2016.06.013

Abstract

Abstract: In order to provide more convenient and efficient restriction enzyme site for the construction of CRISPR/Cas9 gene editing system in the future,the multiple clone site of pBluescript Ⅱ SK(+) and pCAMBⅠA1300 vectors were modified.DNA sequences showed that site-directed mutagenesis were successfully implemented in both pBluescript Ⅱ SK(+) and pCAMBⅠA1300 vectors.The four clone sites of pBluescript Ⅱ SK(+) about Xho Ⅰ, Eco R Ⅴ, Sma Ⅰ, Sac Ⅱ were transformed to Nhe Ⅰ, Mfe Ⅰ, Nsi Ⅰ, Pac Ⅰ respectely;Two clone sites of pCAMBⅠA1300, Sac Ⅰ and Sal Ⅰ were changed to Nsi Ⅰ, Pac Ⅰ.Thus,lay solid foundation for further use on CRISPR/Cas9 genome editing vector.

Key words: pBluescript Ⅱ SK(+), pCAMBⅠA1300, Site-directed mutagenesis, PCR

CLC Number:

PU Yan, LIU Chao, LIN Qi, LI Jiyang, LIU Xiaodong. Site-directed Mutagenesis of pBluescript Ⅱ SK(+) and pCAMBⅠA1300 Vectors Based on PCR Method[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(6): 83-87. doi: 10.7668/hbnxb.2016.06.013.

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