TAILIEUCHUNG - Eukaryotic expression, purification, identification, and tissue distribution of porcine PID1
Phosphotyrosine interaction domain containing 1 (PID1) is a recently discovered gene related to lipid metabolism and may play an important role in fat deposition. In this study, in order to scale up the production of the active recombinant porcine PID1 (pPID1) protein, we reported the expression and purification of a His-tagged version of pPID1 in the yeast Pichia pastoris. | Turkish Journal of Biology Research Article Turk J Biol (2015) 39: 705-711 © TÜBİTAK doi: Eukaryotic expression, purification, identification, and tissue distribution of porcine PID1 Xiaoling CHEN, Huan WANG, Bo ZHOU, Zhiqing HUANG*, Gang JIA, Guangmang LIU, Hua ZHAO Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Chengdu, Sichuan, PR China Received: Accepted/Published Online: Printed: Abstract: Phosphotyrosine interaction domain containing 1 (PID1) is a recently discovered gene related to lipid metabolism and may play an important role in fat deposition. In this study, in order to scale up the production of the active recombinant porcine PID1 (pPID1) protein, we reported the expression and purification of a His-tagged version of pPID1 in the yeast Pichia pastoris. The pPID1 cDNA was cloned into the pPICZαA vector and was expressed in methylotrophic yeast (P. pastoris X33) under control of the alcohol oxidase promoter. The intracellularly expressed recombinant protein was purified by Ni-IDA affinity chromatography, yielding over 95% purity and about mg/L. The recombinant protein was identified by Western blot. In addition, the tissue distribution of pPID1 protein was investigated, and expression of pPID1 protein was mainly detected in the skeletal muscles and liver. This study provides a simple and efficient method for yielding a large amount of active recombinant pPID1, which can be useful for further study of the pPID1 protein. Key words: Porcine PID1, Pichia pastoris, purification, tissue distribution 1. Introduction Phosphotyrosine interaction domain containing 1 (PID1), also known as NYGGF4, was first isolated and characterized from human adipose tissue using suppression subtractive hybridization (Wang et al., 2006). PID1 is primarily expressed in the adipose
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