Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Journal of The Fourth Military Medical University》 2005-22
Add to Favorite Get Latest Update

Expression and purification of recombinant human IL-18 in pichia pastoris

PENG Yan,WANG Yong,SONG Fang-Zhou,WANG Ya-PingKey Laboratory of Biochemistry and Molecular Pharmacology,Chongqing University of Medical Sciences,Chongqing 400016,China  
AIM: To achieve high level expression and purification of recombinant mature human IL-18(mhIL-18)protein in pichia pastoris.METHODS: An Intein Tag was added to the vector.The gene of mhIL-18-Intein was amplified by SOEing and asymmetric PCR.The recombinant expression plasmid pPIC9-IL-18-Intein was constructed by cloning mhIL-18-Intein into pPIC9 vector and was transformed to GS115.The expression of recombinant fusion protein was induced by methanol under an optimum inducing condition.After the protein excreted out of the cells was harvested,SDS-PAGE and Western Blot were used to analyze the expression of recombinant fusion protein.The activity of mhIL-18 purified by affinity chromatography was measured by MTT assays.RESULTS: The cloned sequence of mhIL-18 was identical with the sequence in GenBank.After methanol induction,the fusion protein of mhIL-18 reached the secretion peak of 100 mg/L at 96 h.The purity of the recombinant expressed mhIL-18 was 95% by means of affinity chromatography and the purified mhIL-18 had the biological activity of IL-18.CONCLUSION: We have succeeded in expressing and purifying the recombinant human IL-18 with obvious biological activity in pichia pastoris.
【Fund】: 渝教科2001[12]重庆医科大学重点基金(Cx200101)
【CateGory Index】: R392
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
Chinese Journal Full-text Database 10 Hits
1 LI Wu ping,LV Hong liang,DUAN Zhao jun,ZHANG Li ping,LIU Yong qing,WU Bin wen, CHEN Yong,ZHANG Cheng hai,YI Zhuo an,WEI Kai kun,HOU Yun de (National Institute for Viral Disease Control and Prevention,China CDC,Beijing 10005;High Expression and Purification of IFN-β1b and Its Antiviral Activity[J];Chinese Journal of Virology;2002-03
2 ZHANG Wei CHEN Jin-chun(College of Life Science and Technology,Beijing University of Chemical Technology,Beijing 100029,China);Separation and purification of recombinant snake neurotoxins from Pichia pastoris[J];Journal of Beijing University of Chemical Technology;2005-06
3 CHEN Cui-li~1,SHI Ding-ji~2(1.College of Arts and Science of Beijing Union University,Beijing 100083,China;2.China Institute of Botany,The Chinese Academy of Science, Beijing 100093,China);Construction of Anabaena Transferred the Gene Encoding the Human Granulocyte Colony-stimulating Factor[J];Journal of Beijing Union University;2005-03
4 ZHAO Jing~(*1,2),TANG Huan-wen~1,ZHU Xun-zhi~1(1.Dept.of Appl.Math.,Dalian Univ.of Technol.,Dalian 116024,China;2.Dept.of Math.and Phys.Sci.,Shandong Inst.of Light Ind.,Jinan 250100,China);An improvement of simulated annealing algorithm and study of its application[J];Journal of Dalian University of Technology;2006-05
5 CHEN Zhan-hua~(1),YANG Guang-hui~(*,1,2)(1.Educational College of Jilin Province,Changchun 130020,China; 2.State Key Laboratory of Theoretical and Computational Chemistry,Institute of Theoretical Chemistry,Jilin University,Changchun 130023,China);Study on the three-dimensional structure of rat estrogen sulfotransferase[J];Journal of Molecular Science;2006-05
6 CHEN Yan-Tao, LUO Zhong-Lin, DING Jian-Dong * (Department of Macromolecular Science, Key Laboratory of Molecular Engineering of Polymers of Educational Ministry, Fudan University, Shanghai 200433, China);Preliminary Computer Simulation of Spatial Structure of SARS Coronavirus E-protein[J];Chemical Research In Chinese Universities;2003-08
7 YU Ping (Genetic Engineering Laboratory,Hangzhou University of Commerce,Hangzhou 310035);Advance in Pichia pastoris expression system[J];Industrial Microbiology;2005-03
8 CHEN Hao(School of Mathematics and Computer Science, Hubei University, Wuhan 430062,China);Protein conformation prediction based on simulated annealing algorithm[J];Journal of Hubei University(Natural Science Edition);2005-02
9 LI, Min-Yonga,b LU, Jing-Fenb XIA, Lin*,a (a Department of Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009) (b National Research Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100083);Receptor-based Molecular Modeling Study on Antagonist-Bound Human α_(1A),α_(1B)和α_(1D)-Adrenoceptors[J];Acta Chimica Sinica;2005-20
10 HE, Gua HUANG, Wen-Caib GUO, Li*,a (a Key Laboratory of Drug-Targeting of Education Ministry and Department of Medicinal Chemistry, West China School of Pharmacy, Sichuan University, Chengdu 610041) (b School of Chemical Engineering, Sichuan University, Chengdu 610065);Homology Modeling and Molecular Docking on CCK_1 Receptor[J];Acta Chimica Sinica;2008-01
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved