Construction of hMSH2-deficient cell strain of DNA mismatch repair enzyme

  Objective   To construct hMSH2-deficient cell strain of DNA mismatch repair enzyme which can be used in studying the functions and action mechanisms of hMSH2 gene.

  Methods   The conservative region of hMSH2 gene cDNA was obtained by molecular biology technique.The eukaryotic expression plasmids of hMSH2 gene antisense RNA were constructed through inserting the conservative region fragments into pEGFP-C1 vectors reversedly.HLFs were transfected with the eukaryotic expression plasmids of hMSH2 gene antisense RNA to construct hMSH2-deficient cells(named as "HLFS").The protein expression levels of hMSH2 gene in HLF and HLFS were detected by the western blotting to estimate the effects of antisense inhibition.

  Results   DNA sequencing results was accorded with genbank at the rate of 99.5%, suggesting that the fragment was a part of hMSH2 cDNA.The construction of DNA mismatch repair enzyme hMSH2-deficient cells train was successful and the protein expression level of hMSH2 gene in HLFS was decreased 44% as compared with that in HLF.

  Conclusion   The successes in constructing a hMSH2-deficient strain offered an effective cell strain for studying the function of hMSH2 gene.