Establishment of Mutagenesis Cell Line of Macaca mulatta Gene TRIM5α by TALEN

Abstract

Abstract: Using transcription activator-like effector nuclease（TALEN）to establish the mutagenesis cell line of Macaca mulatta gene TRIM5α is to lay the foundation for further studying the function of TRIM5α gene. The TALEN plasmids targeting TRIM5α was constructed. The site-directed gene mutagenesis donor vector was obtained by site-directed gene mutagenesis technology and the sequence contained the seventh exon of TRIM5α at the position from 1 211 to 1 224. The TALEN plasmids and donor vector were co-transfected into the M. mulatta’s kidney cell line（LLC-MK2）by electroporation. The single cell line was obtained by the infinite dilution method. By extracting the genomic DNA and amplifying the target sequence by PCR，the cell line with base deletion（1 215-1 216）and site mutations（1 213-1 215，1 217）of TRIM5α was screened. Conclusively，the LLC-MK2 single cell line with knockout and site-directed gene mutagenesis of gene TRIM5α was obtained.

Key words: tripartite motif 5 alpha, TALEN, Macaca mulatta, site-directed gene mutagenesis

CLC Number:

10.13560/j.cnki.biotech.bull.1985.2016.12.010

WANG Xiao-li,YU Qing,YUAN Ya-hong,TENG Zhi-ping,LI Dong-sheng,ZENG Yi. Establishment of Mutagenesis Cell Line of Macaca mulatta Gene TRIM5α by TALEN[J]. Biotechnology Bulletin, 2016, 32(12): 53-57.

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