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The Preparation of Gene Vaccine for Egg Drop Syndrome

Author: LiYing
Tutor: LiRongGui
School: Qingdao University
Course: Microbiology
Keywords: Egg drop syndrome virus Gene engineering vaccine Secretary expression Bacillus subtilis
CLC: S858.31
Type: Master's thesis
Year: 2011
Downloads: 32
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Abstract


The DNA encoding C-terminal of fibrin protein of egg drop syndrome virus(EDSV) was amplified by PCR, and then cloned into plasmid pET-15b to construct pET-15b-EDSC. The recombinant plasmid was introduced into E.coli BL21(DE3) to construct engineering bacteria. Recombinant cellulase was successfully expressed in engineering bacterium induced by IPTG as analyzed by SDS-PAGE. The inclusion bodies were gathered by centrifugation, washed and dissolved by urea of 8mol/l. Recombinant protein was purified by Ni2+-NTA resin. The dissoved proteins were dialyzed three times against at 4℃and freeze drying.In order to improve the solubility of recombinant protein expressed in E.coli, the gene encoding C-terminal of fibrin protein of egg drop syndrome virus was cloned into this plasmid to construct pUC18ompA-EDSC. The recombinant plasmid was introduced into E.coli BL21(DE3) to construct engineering bacteria. Recombinant protein could be expressed in engineering bacteria by culturing as analyzed by SDS-PAGE, and partial recombinant protein was secreted into periplasmic space and culture medium. Western blotting analysis indicated the recombinant C-terminal of fibrin protein purified by Ni2+-NTA affinity chromatography could react with antiserum of chicken infected with EDSV, which showed that the recombinant protein was of obvious antigenicity. This study will offer some reference to the production of gene engineering vaccine against EDSV.Gene expression in Bacillus subtilis was also investigated preliminary in the research. The recombinant plasmid was introduced into Bacillus subtilis to construct engineering bacteria. Recombinant protein could be expressed in engineering bacteria by culturing as analyzed by SDS-PAGE, the recombinant EDSV was expressed in other engineering bacteria was further verified.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Poultry > Chicken
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