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Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli
Zhu, Daling; Li, Aihua; Wang, Jianguo; Li, Ming; Cai, Taozhen; Zhu, DL, Wuhan Univ, Chinese Acad Sci, Inst Hydrobiol, Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2007
Source PublicationINDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS
Volume44Issue:4Pages:204-208
AbstractAerolysin is a toxin (protein in nature) secreted by the strains of Aeromonas spp. and plays all important role in the virulence of Aeromonas strains. It has also found several applications such as for detection of glycosylphosphatidylinositol (GPI)-anchored proteins etc. A. hydrophila is a ubiquitous Gram-negative bacterium which causes frequent harm to the aquaculture. To obtain a significant amount of recombinant aerolysin in the active form, in this study, we expressed the aerolysin in E. Coli Under the control of T7 RNase promoter. The coding region (AerA-W) of the aerA gene of A. hydrophila XS91-4-1. excluding partial coding region of the signal peptide was cloned into the vector pET32a and then transformed into E. coli b121. After optimizing the expression conditions, the recombinant protein AerA-W was expressed in a soluble form and purified using His-Bind resin affinity chromatography. Recombinant aerolysin showed hemolytic activity in the agar diffusive hemolysis test. Western blot analysis demonstrated good antigenicity of the recombinant protein.
KeywordAeromonas Hydrophila Cloning Aera Gene Prokaryotic Expression Recombinant Protein Aera-w Hemolytic Activity
DepartmentWuhan Univ, Chinese Acad Sci, Inst Hydrobiol, Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaBiochemistry & Molecular Biology ; Biophysics
Indexed BySCI
Language英语
WOS IDWOS:000250118400002
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Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8398
Collection期刊论文
Corresponding AuthorZhu, DL, Wuhan Univ, Chinese Acad Sci, Inst Hydrobiol, Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Zhu, Daling,Li, Aihua,Wang, Jianguo,et al. Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli[J]. INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS,2007,44(4):204-208.
APA Zhu, Daling,Li, Aihua,Wang, Jianguo,Li, Ming,Cai, Taozhen,&Zhu, DL, Wuhan Univ, Chinese Acad Sci, Inst Hydrobiol, Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2007).Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli.INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS,44(4),204-208.
MLA Zhu, Daling,et al."Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli".INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS 44.4(2007):204-208.
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