Expression,purification and identification of the LBD domain of human PPARα in E. coli

China Biotechnology

研究报告

Expression,purification and identification of the LBD domain of human PPARα in E. coli

Download:

HTML

PDF(0KB) HTML
Export: BibTeX | EndNote (RIS)

Abstract Peroxisome proliferator-activated receptorα(PPARα) is a ligand-activated transcription factor which plays a pivotal role in regulations of metabolism.A cDNA encoding ligand binding domain(LBD）of PPARα was amplified by RT-PCR from human hepatic tissue and the product was inserted into the downstream of the malE gene in the vector pMAL-p2X,which encodes maltose-binding protein (MBP).The recombinant plasmid containing MBP-PPAR gene was transformed into E.coli.TB1.Transformed TB1 was cultured at 37℃ and 200 r/min，and then induced with 0.4 mmol／L IPTG for 6 hours at 30℃ and same r/min. The cells were harvested by centrifugation and broken by sonication.SDS-PAGE analysis showed that the expressed MBP-PPAR fusion protein was soluble and accounted for 31.34％ of the total protein in the supernatant.The MBP-PPARαLBD fusion protein were purified through amylose-resin affinity chromatography and digested by the protease Factor Xa,then separated by Amylose-resin affinity chromatography and DE-52 anion exchange chromatography.The products,MBP-PPARαLBD and PPARαLBD,with high purity were obtained,which provided the necessary material for screening and researching its ligands.

Key words： MBP fusion protein expression Purification PPARαLBD

Received: 16 October 2006 Published: 25 December 2006

	Service
	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

. Expression,purification and identification of the LBD domain of human PPARα in E. coli. China Biotechnology, 2006, 26(12): 18-21.

URL:

https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I12/18

[1]	ZHANG Ling,CAO Xiao-dan,YANG Hai-xu,LI Wen-lei. The Application of Continuous Purification in Affinity Chromatography and Evaluation of Production Scale-up[J]. China Biotechnology, 2021, 41(6): 38-44.

[2]	LV Yi-fan,LI Geng-dong,XUE Nan,LV Guo-liang,SHI Shao-hui,WANG Chun-sheng. Prokaryotic Expression, Purification of LbCpf1 Protein Gene and in Vitro Cleavage Activity Assay[J]. China Biotechnology, 2020, 40(8): 41-48.

[3]	JIANG Dan-dan,WANG Yun-long,LI Yu-lin,Zhang Yi-qing. Study on the Delivery of RGD Modified Virus-Like Particles to ICG Targeted Tumors[J]. China Biotechnology, 2020, 40(7): 22-29.

[4]	XIE Hang-hang,BAI Hong-mei,YE Chao,CHEN Yong-jun,YUAN Ming-cui,MA Yan-bing. The Purification Procedure for the Recombinant HBcAg Virus-like Particle Easy to Generate Aggregation[J]. China Biotechnology, 2020, 40(5): 40-47.

[5]	WEI Wei,CHANG Bao-gen,WANG Ying,LU Fu-ping,LIU Fu-feng. Heterologous Expression, Purification and Aggregation Characterization of Tau Core Fragment 306-378[J]. China Biotechnology, 2020, 40(5): 22-29.

[6]	LIU Zhen-zhen,TIAN Da-yong. Development of Sucrose Density Gradient Centrifugation Purification Process for Rabies Vaccine[J]. China Biotechnology, 2020, 40(4): 25-33.

[7]	ZHU Tong-tong,YANG Lei,LIU Ying-bao,SUN Wen-xiu,ZHANG Xiu-guo. Purification and Crystallization of PcCRN20-C from Phytophthora capsici[J]. China Biotechnology, 2020, 40(1-2): 116-123.

[8]	PAN Bing-jv,ZHANG Wan-yi,SHEN Hui-tao,LIU Ting-ting,LI Zhong-yuan,LUO Xue-gang,SONG Ya-jian. Research Progress on Separation and Purification of Mannan Oligosaccharide[J]. China Biotechnology, 2020, 40(11): 90-95.

[9]	Yu-feng XIE,Xue-mei HAN,Fu-ping LU. Expression, Purification and Enzymatic Properties of β-glucosidase from Lactobacillus paracasei[J]. China Biotechnology, 2019, 39(5): 72-79.

[10]	JING Jia-mei,XUN Xin,WANG Min,PENG Ru-chao,SHI Yi. Expression and Purification of C-terminal of Arenavirus Polymerase and Screening of Crystallization Conditions[J]. China Biotechnology, 2019, 39(12): 18-23.

[11]	ZHU Meng-lu,WANG Xue-yu,LIU Xin,LU Fu-ping,SUN Deng-yue,QIN Hui-min. Heterologous Expression, Purification and Enzymatic Properties of a Novel Leucine 5-Hydroxylase[J]. China Biotechnology, 2019, 39(12): 24-34.

[12]	Chao-di TONG,Jian-ping WU,Li-rong YANG,Gang XU. Crystal Structural Analysis of DehDIV-R by X-ray Crystallography[J]. China Biotechnology, 2018, 38(8): 19-25.

[13]	Jun-jun CHEN,Ying LOU,Yuan-xing ZHANG,Qin LIU,Xiao-hong LIU. *Expression and Purification of Proliferating Cell Nuclear Antigen in Spodoptera frugiperda* Cells**[J]. China Biotechnology, 2018, 38(7): 14-20.

[14]	Shi-jie LI,Yan-kun YANG,Meng LIU,Zhong-hu BAI,Jian JIN. Efficient Expression of SUMO Protease Ulp1 and Used to Express and Purified scFv by His-SUMO tag[J]. China Biotechnology, 2018, 38(3): 51-61.

[15]	Yuan-qiao CHEN,Ding-pei LONG,Xiao-xue DOU,Run QI,Ai-chun ZHAO. Studies on the Protein Purification Ability of an ELP₃₀-Tag in Prokaryotic Expression System[J]. China Biotechnology, 2018, 38(2): 54-60.

Viewed

Full text

Abstract

Cited

Shared

Discussed