title: [Prokaryotic expression of Leptospira interrogans groEL gene and immunoprotection of its products in hamsters].
abstract: To construct a prokaryotic expression system of groEL gene of Leptospira interrogans serogroup Icterohaemorrhagia serovar Lai strain Lai, and to determine the immunoprotective effect of recombinant GroEL protein (rGroEL) in LVG hamsters.
  
  
  
  The groEL gene was amplified by high fidelity PCR and the amplification products were then sequenced. A prokaryotic expression system of groEL gene was constructed using routine genetic engineering technique. SDS-PAGE plus Bio-Rad Gel Image Analyzer was applied to examine the expression and dissolubility of rGroEL protein while Ni-NTA affinity chromatography was used to extract the expressed rGroEL. The immunoprotective rate in rGroEL-immunized LVG hamsters was determined after challenge with L.interrogans strain Lai. The cross agglutination titers of sera from immunized hamsters with different L.interrogans serogroups were detected using MAT.
  
  
  
  The nucleotide and amino acid sequences of the cloned groEL gene were the same as those reported in GenBank. The constructed prokaryotic expression system of groEL gene expressed soluble rGroEL. The immunoprotective rates of 100 and 200 μg rGroEL in LVG hamsters were 50.0 % and 75.0%, respectively. The sera from the rGroEL-immunized LVG hamsters agglutinated all the L.interrogans serogroups tested with different levels.
  
  
  
  The GroEL protein is a genus-specific immunoprotective antigen of L.interrogans and can be used to develop an universal genetically engineering vaccine of Leptospira.
authors.affiliation: Department of Microbiology and Immunology, College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Li
authors.forename: Yinhuan
authors.initials: Y
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.forename: Dongqing
authors.initials: D
authors.lastname: Cheng
mesh_terms: D000372:Agglutination Tests; D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology*; D018834:Chaperonin 60 / Q000235:genetics* / Q000276:immunology; D006224:Cricetinae; D015870:Gene Expression; D007921:Leptospira interrogans / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D018834:Chaperonin 60; D011994:Recombinant Proteins