Validation of the interaction of GALNT14 and MT2A

doi:10.3969/j.issn.1000-1565.2016.01.011

Abstract

Abstract: Co-immunoprecitipation assay and GST pull-down assay were applied to validate the interaction of GALNT14 and MT2A in vivo and in vitro.The cDNA fragment of MT2A was cloned into pET-DsbA and pCMV-Myc.His-Dsba-MT2A and GST-GALNT14 were expressed and purified,and the results of GST pull-down assay showed that MT2A could bound with GALNT14,which confirmed the directly interaction between them in vitro.Then pCMV-Myc-MT2A and pCDNA3.1-Flag-GALNT14 were co-transfected into human breast cancer cell MCF-7.The results of Co-IP illustrated that anti-Myc antibody can precipitated GALNT14 from cell lysates,which revealed MT2A interacted with GALNT14 in vivo.The results showed that GALNT14 could bind MT2A and interact with each other directly in vivo and in vitro,which provided some promising clue for the further exploration into the function of GALNT14.

Key words: N-acetylgalactosaminyltransferase 14(GALNT14), metallothionein 2A(MT2A), GST pull-down assay(GST pull-down), co-immunoprecitipation analysis(Co-IP)

CLC Number:

ZUO Tao,LIU Xuemin,SHAN Jinshuai,TANG Jingzhen,WU Chen. Validation of the interaction of GALNT14 and MT2A[J]. Journal of Hebei University (Natural Science Edition), 2016, 36(1): 65-71.

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