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Codon optimization type 1 type duck hepatitis A virus VP1 gene and application of recombinant protein of codon optimization type 1 type duck hepatitis A virus VP1 gene

A codon-optimized, duck hepatitis A virus technology, applied in the field of veterinary medicine, can solve the problems of affecting the application, affecting the expression of recombinant proteins, and low activity of the expressed protein, and achieves good specificity, high sensitivity, and good intra-batch and inter-batch. repetitive effect

Inactive Publication Date: 2015-11-18
SICHUAN AGRI UNIV
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  • Abstract
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Problems solved by technology

However, when prokaryotic expression of E. coli is used to express proteins, problems such as the partial tropism of codons affecting the expression of recombinant proteins and the low activity of expressed proteins affecting applications are often encountered.

Method used

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  • Codon optimization type 1 type duck hepatitis A virus VP1 gene and application of recombinant protein of codon optimization type 1 type duck hepatitis A virus VP1 gene
  • Codon optimization type 1 type duck hepatitis A virus VP1 gene and application of recombinant protein of codon optimization type 1 type duck hepatitis A virus VP1 gene
  • Codon optimization type 1 type duck hepatitis A virus VP1 gene and application of recombinant protein of codon optimization type 1 type duck hepatitis A virus VP1 gene

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Embodiment Construction

[0034] Preferred embodiments of the present invention are described in detail below. For the experimental methods that do not specify specific conditions in the examples, usually follow the conventional conditions or the conditions suggested by the manufacturer.

[0035] In this example, the codon-optimized VP1 gene was firstly codon-optimized and the codon-optimized VP1 (optiVP1) gene was artificially synthesized, followed by constructing prokaryotic expression recombinant plasmid pET28a(+)-optiVP1, transforming expression bacteria, expressing in large quantities, sonicating, Gel cutting and purification to obtain optiVP1 recombinant protein, and Western blotting analysis confirmed that the protein can have a strong immune reaction with anti-DHAV-1 positive serum, and further determined the coating concentration of optiVP1 recombinant protein and rabbit IgG, gold standard optiVP1 protein and gold standard Goat anti-rabbit IgG working concentration, and then prepared colloidal...

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Abstract

The invention discloses a codon optimization type 1 type duck hepatitis A virus VP1 gene and application of recombinant protein of the codon optimization type 1 type duck hepatitis A virus VP1 gene. A codon optimization type VP1 gene is obtained by optimizing a codon, a colloidal gold strip is prepared from recombinant protein of the codon optimization type VP1 gene and is used for detecting 1 type duck hepatitis A virus antibodies, and a colloidal gold strip which has higher performance and is used for detecting the DHAV-1 antibodies is obtained. The strip is high in sensitivity, and positive serum of DHAV-1 attenuated vaccine immunized ducks can be detected by the strip even after being diluted by 16-32 times. The strip has high specificity. When the 1 type duck hepatitis A virus antibodies are detected through the strip, high intra-assay and inter-assay repeatability is achieved. The prepared strip can be stored for several months under the temperature of 4 DEG C or under the room temperature of 25 DEG C.

Description

technical field [0001] The invention relates to the field of animal medicine, in particular to the application of a codon-optimized VP1 gene and its recombinant protein. Background technique [0002] Duck Viral Hepatitis (Duck Viral Hepatitis) is an acute and highly pathogenic infectious disease of ducklings mainly caused by Duck Hepatitis Avirus type 1 (DHAV-1), with acute onset, rapid spread, short course of disease and high mortality . The clinical manifestations are opisthotonus, hepatomegaly and a large number of hemorrhagic spots in necropsy, which is one of the main diseases that endanger the duck industry. Although chicken embryonized attenuated vaccines and inactivated vaccines for duck hepatitis developed in my country have been applied and promoted throughout the country to immunize breeding ducks and ducklings and have achieved good results, however, the monitoring of DHAV-1 specific antibodies is an important part of evaluation. The key to the immune effect of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/51G01N33/68G01N33/576
Inventor 汪铭书卿莉程安春杨乔陈孝跃
Owner SICHUAN AGRI UNIV
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