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Published on: December 21, 2019
[Expression and antibody preparation of a novel serine protease ESP30]
Ying-ying Zhang1, Fu-yang Li, Wen-hong Zhang
1Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. bioying@fmmu.edu.cn
Aim:
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.
Conclusion:
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.
