Construction and Effect Test of Simple Homologous Recombination Vector pAmpR-NeoR

doi:10.7668/hbnxb.2017.06.004

Abstract

Abstract: In order to construct a donor plasmid that could be used for gene editing in CRISPR/Cas9 system,AmpR-ori fragment and NeoR fragment were amplified with pcDNA3.1(+)-HA -His as template.And then these two fragments were ligated to construct a simple homologous recombination vector pAmpR-NeoR.As cloning sites, BamH Ⅰ and Xho Ⅰ that were introduced on both sides of the NeoR gene were used for the insrtion of the left and right homologous arms.Using this vector,we constructed a donor plasmid for targeting mouse FasL gene.The experimental results showed that the target sequences were genetically edited.Because both BamH Ⅰ and Xho Ⅰ had their own isocaudomers,pAmpR-NeoR had good scalability,flexibility and relative universality for sequence to be clonded,which was easy to further transform as needed to meet the different needs of specific experiments.

Key words: Gene editing, Homologous recombination vector, Isocaudomers

CLC Number:

WANG Yingze, WANG Qiaolan, CAO Qingqing, FU Yu, LIU Chang, WANG Chenchen, DU Chao. Construction and Effect Test of Simple Homologous Recombination Vector pAmpR-NeoR[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 25-30. doi: 10.7668/hbnxb.2017.06.004.

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http://www.hbnxb.net/EN/Y2017/V32/I6/25

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