Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Journal of Anhui Agricultural Sciences》 2009-04
Add to Favorite Get Latest Update

Cloning Expression and Purification of Heat Shock Protein HSP60 from Hyperthermophilic Archaeon Pyrococcus furiosus

CHEN Hua-you et al(Institute of Biological Sciences,Jiangsu University,Zhenjiang,Jiangsu 212013)  
[Objective] The aim was to study the cloning,expression and purification method of heat shock protein HSP60 from Hyperthermophilic Archaeon Pyrococcus furiosus.[Method] The homology analysis of Pfu-HSP60 gene sequence was taken by using Canada HSP60 database and sequence comparative tools of BLAST and FASTA.HSP60 gene was amplified in vitro and cloned and expressed in Escherichia coli.Pfu-HSP60 protein was also purified.[Result] With the Taq enzyme as DNA polymerase,the anticipative Pfu-HSP60 gene was amplified by PCR method,then it was inserted in plasmid pET-21a and transformed into E.coli BL21-Codonplus(DE)3-RIL.The soluble high expressive protein was obtained after induced by 0.1 mmol/LIPTG for 2 h.After ultrasonic and centrifugation,the supernate was heated at 85 ℃ and centrifuged again,then the primary purified Pfu-HSP60 protein was obtained and then it was purified by DEAE Sepharose Fast Flow anion exchange chromatography.After the collected peak was filtered and concentrated,the Pfu-HSP60 protein peak was collected through Sephacryl S-200 gel filtration column.The purity of the recombinant protein Pfu-HSP60 was 94.5% and the total yield was 28.0%.[Conclusion] The research laid the foundation for the study of the structure and function of Pfu-HSP60 protein and the study of the interaction mechanism with other HSP.
【Fund】: 国家基础研究973计划(2004CB719606);; 江苏大学校基金资助项目(08jdg011)资助
【CateGory Index】: Q93;Q78
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
【Citations】
Chinese Journal Full-text Database 2 Hits
1 ZHOU Mei-Xian XIANG Hua+* TAN Hua-Rong(Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China);Development of the Genetic Transformation System in Extremely Halophilic Archaea[J];Chinese Journal of Biotechnology;2002-03
2 XU Xue-wei~1, WU Min~1, HUANG Wei-da~2 (1.College of Life Sciences, Zhejiang University, Hangzhou 310027, China; 2.College of Life Sciences, Fudan University, Shanghai 200433, China);Isolation and 16S rDNA sequence analysis of extremely halophilic archaea from Saline in Altun region[J];Journal of Zhejiang University(Engineering Science);2004-04
【Co-citations】
Chinese Journal Full-text Database 7 Hits
1 WEN Hong-yu et al(School of Environmental Science and Spatial Informatics,China University of Mining and Technology,Xuzhou,Jiangsu 221116);Culture Condition Optimization of Halophilic Bacteria HBCC-3 and Its Phylogenetic Analysis[J];Journal of Anhui Agricultural Sciences;2008-14
2 WEN Hong-yu1,3,LV Ai-Jun3,HAN Bao-ping1,2 (1.School of Environmental Science and Spatial Informatics,China University of Ming and Technology,Xuzhou Jiangsu,221008;2.Xuzhou Normal University,Xuzhou Jiangsu 221116;3.School of Life Sciences,Xuzhou Normal University,Xuzhou Jiangsu 221116);Culture characteristics and analysis of halophilic archaea strain HBCC-2 by 16S rRNA gene sequence analysis[J];Industrial Microbiology;2008-06
3 ZHOU Xu- hua1,2, WU Min2 (1. Department of Agronomy, Changzhi Vocational and Technical College, Changzhi, Shanxi 046000; 2. College of Life Sciences, Zhejiang University, Hangzhou Zhejiang 310027, China);Phylogenetic Analysis of Twenty-six Halophiles from Zhoushan Islands[J];Modern Agricultural Sciences;2009-04
4 MAO Pei-Hong~(1,2),JIN Xiang~2,WANG Yun~1,IDU Kai~(1*) (1.The Key Lab of Microorganisms in Xinjiang Specific Environment,Ururnqi 830000,China;2.The Center of Ion Beam Biotechnology,Xinjiang University,Urumqi 830008,China);Research and Development on the Resources of Microorganisms in Xinjiang Specific Environment[J];Biotechnology;2006-05
5 TIAN Xin-Peng ZHANG Yu-Qin TANG Shu-Kun LI Wen-Jun ** XU Li-Hua JIANG Cheng-Lin(Laboratory for Conservation and Utilization of Bio-resources,Yunnan Institute of Microbiology, Yunnan University,Kunming 650091);Culture-dependent Extremely Halophilic Archaea Isolated from Heijing Ancient Salt Mine,Yunnan[J];Microbiology;2006-06
6 XU Xiao-Hong1,WU Min2,Zhang Hui-Bin3,LIU Zhi-Hu3 1.Ningbo Institute of Technology,Zhejiang University,Ningbo 315100,China;2.College of Life Science,Zhejiang University,Hangzhou 310027,China;3.The Altun Mountain National Nature Reserve,Kuerle 841000,China;Genetic analysis of the br gene in halophilic archaea isolated from Xinjiang region[J];Hereditas;2007-03
7 Xugela·HABUDIN~1,CHEN Jin~1,Dliwuzi·ANIWAR~2 HUANG Ruihu~1,ZHOU Yuguang~3,Dlibar·TUOHUTI~1 (1.College of Life Science and Chemistry,Xinjiang Normal university,Xinjiang Urumqi 830054; 2.College of Life Science,Xinjiang University,Urumqi Xinjiang 830046);Studies on Physiological and Biochemical Characters and Bacteriorhodopsin Gene of Strain ABDH12 of XingjiangAibi Lake[J];Journal of Xinjiang Normal University(Natural Sciences Edition);2009-04
【Secondary Citations】
Chinese Journal Full-text Database 3 Hits
1 XU Xue-wei 1), WU Min 1)*, RUAN Hong 1), HONG Jie 2), HUANG Wei-da 2) ( 1)College of Life Sciences, Zhejiang University, Hangzhou 310027, China; 2)College of Life Sciences, Fudan University, Shanghai 200433, China);Partial Sequence of the Gene for a New BR Protein, and Nucleotide Sequence of 16S rRNA Encoding Gene from a Halophilic Archaeum AJ4[J];Chinese Journal of Biochemistry and Molecular Biology;2004-01
2 Chen Wenxin(College of Biological Sciences,China Agricultural Universyty,Beiji 100094);BACTERIAL PHYLOGENY[J];ACTA MICROBIOLOGICA SINICA;1998-03
3 LI Ling\+1, WU Min\+1, QIAO Shou\|yi\+2, CHEN Kang\|sheng\+3\;(1.Life Science College, Zhejiang Uni., Hangzhou 310027, China; 2.Genetics Institution, Fudan Uni., Shanghai 200433, China; 3.Dept. of Information & Electronic Engineering, Zhejiang Uni., Han;Cloning and expression of the Bacteriorhodopsin gene[J];Journal of Zhejiang University(Engineering Science);2001-03
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved