Eukaryotic expression, purification, identification, and tissuedistribution of porcine PID1

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.

📌 3 Maddede Bu Makale

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Amaç & Hipotez: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.

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Temel Bulgular & Veri: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.

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Sonuç & Katkı: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.

🔬Tür:Araştırma Makalesi

📍Kapsam:Genişletilmiş Akademik Örneklem

⚙️Değişkenler:Porcine PID1 · Pichia pastoris · purification

Özet

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 1.8 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.

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 1.8 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.

Anahtar kelimeler

Porcine PID1 Pichia pastoris purification tissue distribution

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Bu Makaleye Atıf Yap

CHEN, X., WANG, H., ZHOU, B., HUANG, Z., JIA, G., LIU, G., ZHAO, H. (2015). Eukaryotic expression, purification, identification, and tissuedistribution of porcine PID1. Turkish Journal of Biology, Cilt: 39 - Sayı: 5, 705–711. https://ajindex.com/articles/181476

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Kaynakça (9)

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