Expression and antibody preparation of a novel serine protease ESP30

Ying ying Zhang, Fu yang Li, Wen hong Zhang, Jian Zhang, Yong Huang, Xin ping Liu, Li bo Yao

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)334-336, 339
JournalXi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Volume21
Issue number3
StatePublished - May 2005

ASJC Scopus subject areas

  • Medicine(all)

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  • Cite this

    Zhang, Y. Y., Li, F. Y., Zhang, W. H., Zhang, J., Huang, Y., Liu, X. P., & Yao, L. B. (2005). Expression and antibody preparation of a novel serine protease ESP30. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 21(3), 334-336, 339.