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Classic swine fever virus (CSFV) C strain E2 truncated protein and its preparation method and use

A technology of swine fever virus and truncated protein, which is applied in the direction of biochemical equipment and methods, viruses, virus peptides, etc., and can solve problems such as artificiality and uncertainty

Active Publication Date: 2014-02-19
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The value measured by the traditional ELISA method at the end of color development will change with time, such a situation will lead to obvious artificial and uncertain results

Method used

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  • Classic swine fever virus (CSFV) C strain E2 truncated protein and its preparation method and use
  • Classic swine fever virus (CSFV) C strain E2 truncated protein and its preparation method and use
  • Classic swine fever virus (CSFV) C strain E2 truncated protein and its preparation method and use

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Embodiment 1

[0080] ⅠMaterial

[0081] The RNA extraction kit was purchased from OMEGA; the KOD FX enzyme was produced by TOYOBO;

[0082] The PCR cleaning kit was purchased from SIMGEN; the PCR recovery kit was purchased from SIGMA;

[0083] The prokaryotic expression vector pET-32a was purchased from Novagen;

[0084] Restriction enzymes BamH Ⅰ, Hind Ⅲ, T4 DNA ligase, DNA Marker, protein Marker, Escherichia coli DH5α competent cells, Escherichia coli BL21 competent cells, etc. were purchased from TaKaRa Company;

[0085] His-TRAP TM FF Crude purification column was purchased from GE Healthcare;

[0086] The attenuated swine fever rabbit vaccine was purchased from Guangdong Yongshun Biopharmaceutical Co., Ltd.;

[0087] The ELISA standard kit was purchased from IDEXX Company, and the standard serum was referred to the negative and positive pig serum in the ELISA kit of IDEXX Company.

[0088] Ⅱ specific preparation process

[0089] One, the preparation of the CSFV E2 truncated prot...

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Abstract

The invention discloses a classic swine fever virus (CSFV) C strain E2 truncated protein and its preparation method and use. The main antigen region of the E2 gene is expressed and three pairs of disulfide bonds for maintaining the antigen space structure are expressed simultaneously so that the expressed protein has a certain space structure and satisfies natural protein characteristics and the inclusion body is conducive to protein purification. The E2 truncated protein obtained by the invention is used for building a CSFV IgG antibody ELISA detection method. A result shows that the recombinant protein has good antigenicity. The built ELISA kit adopting the CSFV C strain E2 truncated protein has good specificity, accuracy, sensitivity and repeatability after use. A substrate coloration solution used in the invention has an indication effect, and after several hours to 4 nights after the reaction, the detection result still has credibility. Therefore, the coloration system can reduced human factor influences, avoids false positive appearance, makes a negative result stable and guarantees good detection on CSFV antibodies.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a truncated protein of swine fever virus C strain E2, a preparation method and application thereof. Background technique [0002] Classical Swine Fever (CSF) is an important viral infectious disease that seriously endangers the pig industry. The disease is prevalent worldwide. It is a major severe infectious disease of Class A stipulated by OIE, and its pathogen is also listed in the International Biological Weapons Convention. Listed as an important animal biological warfare agent or animal bioterrorism pathogen, it is highly valued by the international community. [0003] Swine fever is an acute, febrile, highly contagious infectious disease of pigs caused by swine fever virus, also known as swine cholera. Classic swine fever virus (CSFV, formerly known as Hog cholera virus, HCV) is now a member of Flaviviridae and Pestivirus, members of the same genus also include bov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/18C12N15/70G01N33/68G01N33/569
CPCC07K14/005C12N15/70C12N2770/24322G01N33/56983G01N2333/183G01N2469/20
Inventor 陈金顶郑仲华姚俊庸李东升勾红潮漆月王利军邓洁汝
Owner SOUTH CHINA AGRI UNIV
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