title: [Expression and antibody preparation of a novel serine protease ESP30].
abstract: OBJECTIVE
  To express ESP30, a novel serine protease, in Escherichia coli and to prepare anti-ESP30 antibody.
  
  
  METHODS
  ESP30 gene was amplified by PCR from the genome of aeromonas hydrophila, and cloned into the expression vector pDH2. The ESP30 expression was carried out under thermal induction. The antiserum was prepared by immunizing rabbit with ESP30. The titer and specificity of the antibody were detected by ELISA and Western blot respectively.
  
  
  RESULTS
  The ESP30 non-fusion protein with relative molecular mass (M(r)) of 66,000 was highly expressed in E. coli. The rabbit antibody against ESP30 was obtained. The ELISA titer of antiserum against ESP30 was about 1:128,000. Western blot analysis showed that the antiserum could bind to the expressed ESP30 specifically.
  
  
  CONCLUSIONS
  The rabbit antibody against ESP30 has been successfully prepared, which lays the foundation for further studying the structure and function of the novel protease ESP30.
authors.affiliation: Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. bioying@fmmu.edu.cn
authors.forename: Ying-ying
authors.initials: YY
authors.lastname: Zhang
authors.forename: Fu-yang
authors.initials: FY
authors.lastname: Li
authors.forename: Wen-hong
authors.initials: WH
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Xin-ping
authors.initials: XP
authors.lastname: Liu
authors.forename: Li-bo
authors.initials: LB
authors.lastname: Yao
mesh_terms: D016980:Aeromonas hydrophila / Q000201:enzymology*; D000906:Antibodies / Q000032:analysis / Q000276:immunology*; D000918:Antibody Specificity; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D012995:Solubility
chemical_list: D000906:Antibodies; D012697:Serine Endopeptidases