IMR Press / FBE / Volume 2 / Issue 3 / DOI: 10.2741/E172

Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on as a courtesy and upon agreement with Frontiers in Bioscience.

Prokaryotic expression and characterization of human AC3-33 protein
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1 Department of Life Science, North China Coal medical college, No. 57 JianShe South Road, Tangshan 063000, China
2 State Key Laboratory of Animal Nutrition, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, China
3 School of Biology & Food Technology, Dalian Polytechnic University, Dalian 116034, China
4 Department of Environment & Chemical Engineering, Tangshan College, Tangshan 063000, China
Front. Biosci. (Elite Ed) 2010, 2(3), 1134–1142;
Published: 1 June 2010

The transcription factor, AP-1, plays an important role in cellular proliferation, transformation and death. We previously showed that AC3-33 (GenBank name: c3orf33, FLJ31139), significantly inhibited transcriptional activity of AP-1. In this study, we report a method to express and purify AC3-33 in E. coli using glutathione-S-transferase (GST) fusion system. A GST-fusion protein was created by insertion of AC3-33 gene into a pGEX-4T-1 vector. The fusion protein, GST-AC3-33, was expressed in BL21 strain, and purified by GSH-affinity chromatography followed by thrombin cleavage. The digested product was further purified in a GSH-affinity column. After cleavage and purification, the recombinant AC3-33 protein exhibited the expected size of 29 kDa by SDS-PAGE and Western blotting and inhibited transcriptional activity of AP-1 in a dual-luciferase reporter assay. The bioactive recombinant GST-AC3-33, can be used to decipher the physiological and biochemical role of this protein.

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