Human NDRG1 of the GST fusion expression purification and detection of interacting proteins.docVIP

 PAGE \* MERGEFORMAT 11 Human NDRG1 of the GST fusion expression purification and detection of interacting proteins Author: Qian Yang, Zhang, Zhang Yingying, He Peng, Liu Xinping, Yao Libo, Chiu Wai-yin [Keywords:] glutathione agarose GST fusion expression and purification of human NDRG1 [Abstract] AIM: To better understand the function of NDRG1 and look for the interact protein through pulldown technic. METHODS: We digested the NDRG1 cDNA from the constructed vector NDRG1pPROEXHTb and transfered it into another vector pGEX4T1. After gene was sequenced, the recombinant vector NDRG1pGEX4T1 was transformed into E. coil DH5α and the strains highly expressing soluble GSTNDRG1 in LB medium containing ampicillin were obtained. The fusion protein was then purified by Glutathione Sepharose 4B and was incubated with HHCC. The interact protein with NDRG1 was observed by SDSPAGE. RESULTS : There was a new strip at Mr 4.0 * 104 when the purified protein and HHCC were incubated. CONCLUSION: The interact protein with NDRG1 is found in HHCC. [Keywords] NDRG1; GST; gene expression; glutathione sepharose 4B [Abstract] Objective: To better understand the function of NDRG1 through the pulldown technique to search for proteins that interact with NDRG1. Methods: Construction of a good NDRG1pPROEXHTb from a recombinant vector digestion under NDRG1 cDNA, into fusion expression vector pGEX4T1, Sequencing later transformed into DH5α competent cells, in LB containing ampicillin in the highly expressed. GSTNDRG1 through the expression of glutathione agarose 4B affinity chromatography purified, and with high expression cell line HHCC incubated with each other by SDSPAGE detection and NDRG1 the role of the protein. Results: The purified protein and incubated HHCC after Mr 4.0 * 104 occurring at the new bands should be the unknown proteins interact with NDRG1. Conclusion: There HHCC in tumor cells interact with NDRG1 protein. [Keywords:] NDRG1; GST; gene expression; glutathione-S