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Segmented Expression of Structural Protein of Foot-and-Mouth Disease Virus and Immunization Test

Author: MaXiaoXia
Tutor: ShenXinLiang
School: Lanzhou University
Course: Biochemistry and Molecular Biology
Keywords: FMDV structural protein protein expression antigenic characteristics sub-unit vaccine
CLC: S852.65
Type: Master's thesis
Year: 2009
Downloads: 90
Quote: 1
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Abstract


Foot-and-mouth disease virus (FMDV) is the causative agent of the economically most important animal virus disease world-wide .Although mortality associated with FMD is usually low, the disease decreased livestork productivity, and affected contries cannot participate in international trade of animals and products . With applications of molecular biology technique to FMDV, FMDV toxic proteases have been research hot point. Moreover, primal antigen site gene of FMDV was expressed in prokaryotic cell and product was purified for inoculating animal. This study will lay a foundation for understanding sub-unit vaccine development.Foot-and-mouth disease virus RNA was used as template for RT-PCR. The amplified cDNA products were cloned into pGEM-T Easy Vectors and expression Vectors. By means of DNA star, we can predict and analyse the antigen sites of the three proteins. At the same time we optimized the condition of expression system. Later, we confirmed the form of expressing protein is inclusion body. We purified the inclusion by using nickel chelate affinity chromatography. Then, after dialysis renaturation and concentration, we injected the ptoteins into the mouse .It is indicated that the proteins have antigenic characteristics and can induce antibodies.The proteins were tested by antibody sandwiched ELISA, and the results showed that all of the OD values range from the OD value of positive control to that of negative control, and the VP1 sample was the highest in those tested samples. The result showed that according to the ability of inducing antibodies, VP1 had a stronger function than the rest. The complexes, including VP1 and VP0, or VP1 and VP3, or VP0 and VP3, or VP0, VP1 and VP3, failed to overcome the VP1 at the aspect of the ability to induce antibody.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Livestock Virology
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