关键词: 小鼠, 基因敲除, 打靶载体

Abstract: Objective To amplify the enamel matrix serine proteins 1(EMSP1) in vitro and to establish gene targeting vector and transgenic mouse models to study the physiological and pathological function of EMSP1.Methods Two pairs of primers were d esigned by Oligo 6.0 software and synthesized according to EMSP1 gene sequence. Two gene fragments of 129 strain mouse EMSP1 were amplified from mouse genomic DNA by PCR-5 000 bp 5′ arm and 1 700 bp 3′arm. The two arms were cloned into a universal gene knockout vector-pSSC-9 respectively on either side of the positive selective marker neo. The targeting vector was identified by PCR， restriction endonuclease and sequence analysis. Results The restriction endonuclease identifiation result indicated that 5 000 bp and 1 700 bp fragments were obtained and homogeneous long and short arms were cloned into vector.DNA squence analysis showed that EMSP1 gene targeting vector pSSC-9-EMSP1 was successfully constructed. Conclusion The mouse EMSP1 gene targeting vector is constructed successfully.

Key words: mice gene knockout, targeting vector