Membrane protein gene of chicken infective bronchitis virus and clone method thereof

A bronchitis, chicken infectious technology, applied in the direction of genetic engineering, plant genetic improvement, chemical instruments and methods, etc.

Inactive Publication Date: 2009-10-21
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the research on the membrane protein gene of chicken infectious bronchitis virus at home and abroad is still inconclusive. Therefore, the systematic and in-depth research on the membrane protein gene of chicken infectious bronchitis virus will reveal the prevalence and variation of IBV from the molecular level , it is very necessary to prevent and control the occurrence and prevalence of IB in my country

Method used

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  • Membrane protein gene of chicken infective bronchitis virus and clone method thereof
  • Membrane protein gene of chicken infective bronchitis virus and clone method thereof
  • Membrane protein gene of chicken infective bronchitis virus and clone method thereof

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Embodiment Construction

[0022] The present invention will be described in detail below with reference to the drawings and specific embodiments.

[0023] 1 material

[0024] 1.1 Strain

[0025] The IBV-HN99 strain has been applied for a Chinese patent (patent application number is 200610018065.4), and it was published on January 9, 2008.

[0026] 1.2 SPF chicken embryo

[0027] The 9-11-day-old SPF chicken embryos were provided by Zhengzhou Biopharmaceutical Factory of China Animal Husbandry Co., Ltd.

[0028] 1.3 Vectors and strains

[0029] The cloning vector pMD18-T is a product of TaKaRa; Escherichia coli TOP10 was purchased from the State Key Laboratory of Medical Molecular Virology, Wuhan University.

[0030] 1.4 Tool enzymes and reagents

[0031] All restriction enzymes used were purchased from TaKaRa; Trizol RNA extraction kit was a product of Invitrogen; Reverse Transcription System was a product of ToYoBo; DNA rapid recovery kit was purchased from Shanghai Shenneng Bioengineering Co., Ltd.; rapi...

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Abstract

The invention discloses a membrane protein gene of an infective bronchitis virus of chicken. A complete sequence of the gene comprises 669 ribotides for coding M proteins of 222 amino acid residues. The discoveries of a molecule genetic variation rule of the membrane protein gene of the chicken infective bronchitis virus and an epitope on the coded membrane protein of the gene have important significance to the development and the application of infective bronchitis genetic engineering antigens and genetic engineering vaccines. The invention also discloses a method for cloning the gene, wherein 5'-CGGAATTCAGTTTCCTAAGAACGGTTGGAA-3' is used as an upper primer and 5'-CCCAAGCTTCTCTCTACACGCACACATTTAT-3' is used as a lower primer for amplifying a full-length segment of the membrane protein gene of a chicken infective bronchitis virus HN99 through an RT-PCR method.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a chicken infectious bronchitis virus membrane protein gene and a cloning method thereof. Background technique [0002] Infectious Bronchitis (IB) is an acute and highly contact infectious disease of chickens caused by Infectious Bronchitis Virus (IBV), which mainly affects the respiratory, digestive and genitourinary systems of chickens. , Resulting in a decrease in the number of laying hens and lower egg quality; chicks died in large numbers due to respiratory or kidney infections. In addition, the disease can also cause chickens to gain weight and reduce feed returns, causing significant economic losses to the poultry industry, and becoming one of the most important infectious diseases hindering the development of the world's poultry industry. [0003] IBV is a representative species of the Coronavirus family and the genus Coronavirus. It has an outer envelope and is a medium pleomorph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/50C12N15/10C07K14/165
Inventor 贺秀媛张淑霞李玉峰杜恩岐崔沛张君涛张志平崔保安杨明凡陈红英邓立新贺丛董海聚仝宗喜张惠茹白延杰安森亚朱翠娟原维李小波
Owner HENAN AGRICULTURAL UNIVERSITY
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