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:: Volume 14, Issue 3 (7-2015) ::
IJFS 2015, 14 Back to browse issues page
Prokaryotic expression, purification and immunogenicity analysis of CpsD protein from Streptococcus iniae
H.C. Wang , K.Y. Wang * , J Wang , D Xiao , J.L. Huang , D.F. Chen
, kywangsicau@126.com
Abstract:   (2809 Views)
Streptococcus iniae is a major cause of serious bacterial infections in both fish and human beings. Capsular polysaccharide (CPS) of S. iniae is vital to evade phagocytic clearance of the host and serves as an important protective antigen of S. iniae infection in aquatic animals. The CpsD gene was determined to be highly conservative in capsule polysaccharide operon. Prokaryotic expression of the CpsD gene of a clinical isolate of S. iniae from channel catfish and immunogenic examination of the recombinant protein were first described in this essay. The recombinant protein was expressed in the form of inclusion bodies (IBs). Induction conditions in Escherichia coli were optimized with 0.6mM Isopropyl β-D-1-Thiogalactopyranoside at 37°C for 5h after the culture mid-log phase in Luria Bertani (LB) medium. The recombinant protein CpsD was thus expressed and purified by immobilized metal affinity chromatography (IMAC), yielding approximate 582.47 mg the protein per liter culture. Western blot analysis showed that the purified CpsD had reactogenicity. It will possibly reveal more details of capsule synthesis and capsule regulation during various stages of the S. iniae infectious process.
Keywords: Streptococcus iniae, Capsular polysaccharide, Prokaryotic expression, Purification, Western blot analysis.
Full-Text [PDF 1045 kb]   (939 Downloads)    
Type of Study: Orginal research papers | Subject: fish disease
Received: 2013/02/19 | Accepted: 2014/09/17 | Published: 2015/08/4
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Wang H, Wang K, Wang J, Xiao D, Huang J, Chen D. Prokaryotic expression, purification and immunogenicity analysis of CpsD protein from Streptococcus iniae . IJFS. 2015; 14 (3) :612-622
URL: http://jifro.ir/article-1-903-en.html


Volume 14, Issue 3 (7-2015) Back to browse issues page
مجله علوم شیلاتی ایران Iranian Journal of Fisheries Sciences
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