Cloning, Expression, and Immunogenicity Analysis of the CpsE Protein from Group B Streptococcus Isolated from Tilapia Oreochromis NiloticusReport as inadecuate




Cloning, Expression, and Immunogenicity Analysis of the CpsE Protein from Group B Streptococcus Isolated from Tilapia Oreochromis Niloticus - Download this document for free, or read online. Document in PDF available to download.

Group B Streptococcus GBS is amajor cause of serious bacterial infection in numerous animal species. Theproduction of capsular polysaccharideCPs is vital to GBS to evade hostimmunity. One of the genes that required for production of CPs, cpsE, has beendetermined to be well conserved in capsule gene cluster cps.This study clonedthe cpsE gene from Tilapia of GBS clinical isolate serotype Ia and expressedthis gene with aid of pET-32a+ in Escherichia coli BL21DE3 competent cellsto obtain high levels of the recombinant protein for further study about CpsEin fish and examination of its immunogenicity. The optimization of inductionconditions IPTG concentration, temperature and time in E.coli was accomplishedand let us to perform the recombinant protein induction at 37℃ for 3h,with 0.2mM IPTG in Luria Bertani LB medium.At the optimal conditions, recombinant protein was expressed in an insolubleform inclusion bodies and accounted for approximately 23% of the totalprotein. Purification by affinity chromatography yielded about 480mg fusionprotein per liter culture.

KEYWORDS

Recombinant CpsE Expression; Purification; Polyclonal Antibody

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X. Fu, K. Wang, J. Huang, J. Wang, H. Lian, Y. He, L. Li and K. Wang -Cloning, Expression, and Immunogenicity Analysis of the CpsE Protein from Group B Streptococcus Isolated from Tilapia Oreochromis Niloticus,- Engineering, Vol. 4 No. 10B, 2012, pp. 167-171. doi: 10.4236-eng.2012.410B044.





Author: Xi Fu, Kaiyu Wang, Jinlu Huang, Jun Wang, Hai Lian, Yang He, Lanmin Li, Kaiyu Wang

Source: http://www.scirp.org/



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