[Expression and antibody preparation of a novel serine protease ESP30].
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.
Zhang
Ying-ying
YY
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. bioying@fmmu.edu.cn
Li
Fu-yang
FY
Zhang
Wen-hong
WH
Zhang
Jian
J
Huang
Yong
Y
Liu
Xin-ping
XP
Yao
Li-bo
LB
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
D000906:Antibodies; D012697:Serine Endopeptidases