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Antigen protein and encoding gene and applications thereof

A technology of antigenic protein and gene, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of biological safety risk, inability to preserve for a long time, poor sensitivity, etc., and achieve high sensitivity and specificity, high specificity and sensitivity , the effect of less sample demand

Inactive Publication Date: 2011-09-07
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
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  • Claims
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Problems solved by technology

However, there are also some practical problems, such as: (1) the source and collection of human "O" red blood cells are inconvenient, and they cannot be stored for a long time, which limits the clinical application of this method; (2) the judgment of the results is easily affected by subjective factors influence, prone to deviation
For example, Italian and German scholars found that HA can produce 8% false positive or false negative by comparing HA and ELISA, and the sensitivity is poor; (3) recent studies have found that some RHDV isolates do not produce hemagglutination , so the HI or HA method cannot be used for detection; (4) HA and HI methods use highly infectious RHDV, which brings potential biosafety risks to the operator and the surrounding environment

Method used

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  • Antigen protein and encoding gene and applications thereof
  • Antigen protein and encoding gene and applications thereof
  • Antigen protein and encoding gene and applications thereof

Examples

Experimental program
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Embodiment Construction

[0026] obtain target protein

[0027] Liver tissues of dead rabbits infected with rabbit haemorrhagic disease virus RHDV JX97 (full genome sequence GENBANK accession number DQ205345) were collected, and viral RNA was extracted with UNIQ-10 Column TRIzol Total RNA Extraction Kit (Shanghai Sangong). The specific operation was carried out according to the instructions. Use the extracted total viral RNA as a template for reverse transcription, and 20 μL of the reverse transcription system is as follows:

[0028] Viral RNA 10μL; 5×M-MLV Buffer 4μL, dNTP (10mmol / L) 1μL; RNase inhibitor (20U / L) 0.5μL; M-MLV 1μL VP60 gene downstream primer (5'-GCAAGTCCCAATCCGATGAAT-3') 1μL,; DEPC Make up the volume to 20 μL.

[0029] The reaction conditions are as follows:

[0030] Water bath at 70°C for 10 minutes, transfer to a constant temperature water bath at 42°C for 1 hour, and then cool in ice for 2 minutes to obtain the first strand of VP60 cDNA.

[0031] Using VP60cDNA as a template for P...

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Abstract

The invention discloses an antigen protein and encoding gene and applications thereof. The amino acid sequence of the antigen protein is shown in the SEQ ID NO.1. The antigen protein relates to deficiency virus antigen, has good safety and does not contain unrelated impurity protein; the antigen protein is only bound with the antibody of the rabbit hemorrhagic disease virus (RHDV) in serum and can not perform cross reaction with the positive serums of other rabbit diseases, thus the antigen protein has higher sensitivity and specificity. By adopting the genetic engineering means, the antigen protein is obtained through cloning and expression; and in the expression process, the antigen protein is accumulated in the host strain in an inclusion mode and is easy to separate and purify, thus the industrial production can be performed.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to an antigenic protein and its encoding gene and application. Background technique [0002] Rabbit hemorrhagic disease (RHD), commonly known as "rabbit plague", is an acute and highly fatal rabbit infectious disease caused by rabbit hemorrhagic disease virus (RHDV). The disease has the characteristics of high fatality rate, fast death, and rapid spread, which poses a serious threat to the healthy development of the rabbit industry. Therefore, it is officially listed as a B-type infectious disease in the "International Animal Health Catalog" by the International Bureau of Animal Health. The Ministry of Agriculture of my country It is also listed as a second-class animal disease. Rabbit viral hemorrhagic disease was first discovered and reported by Chinese scholars in 1984. Subsequently, major rabbit-raising countries in the world, such as Europe, Oceania, North Africa, Australia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/08C12N15/70C12N15/40G01N33/569C12R1/19
Inventor 余斌刘光清云涛倪征陈柳华炯钢李双茂梁华丽
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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