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Construction method of cow mastitis pathogenic bacterium triple epitope gene and garrupa c type lysozyme gene recombinant adenovirus and application

A cow mastitis and construction method technology, which is applied in the field of construction and application of recombinant adenovirus with triple epitope gene of cow mastitis pathogenic bacteria and c-type lysozyme gene of grouper, can solve bacterial drug resistance and drug residue And other issues

Active Publication Date: 2017-12-19
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

The frequent use of antibiotics can easily lead to problems such as bacterial resistance and drug residues, and it is not a long-term treatment method that can be relied on

Method used

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  • Construction method of cow mastitis pathogenic bacterium triple epitope gene and garrupa c type lysozyme gene recombinant adenovirus and application
  • Construction method of cow mastitis pathogenic bacterium triple epitope gene and garrupa c type lysozyme gene recombinant adenovirus and application
  • Construction method of cow mastitis pathogenic bacterium triple epitope gene and garrupa c type lysozyme gene recombinant adenovirus and application

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Embodiment Construction

[0027] Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail:

[0028]Cow mastitis is mainly caused by the cross-infection or co-infection of three pathogenic bacteria, Staphylococcus aureus, Escherichia coli and Streptococcus. At this stage, antibiotics are mainly used as prevention and treatment methods . The frequent use of antibiotics can easily lead to problems such as bacterial resistance and drug residues, and it is not a long-term treatment method that can be relied on. Epitope vaccines have the advantages of good safety, strong protection, and adjustable response types. They can activate specific immune cells in the body to respond by continuously expressing antigenic substance epitope polypeptides in cells. In the experiment, three specific proteins of the main pathogenic bacteria of dairy cow mastitis were selected, namely, Ebps and ClfA of Staphylococcus aureus, OmpA and OmpC of Escherichia coli...

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Abstract

The invention discloses a construction method of a cow mastitis pathogenic bacterium triple epitope gene and garrupa c type lysozyme gene recombinant adenovirus and application. In the tests, cow mastitis main pathogenic bacterium special proteins are selected, two B cell epitopes of each protein are obtained through prediction of an ABCpred scheme and a BepiPred scheme and are connected to obtain a TRE sequence, a cLYZ sequence is obtained on GeneBank, and a mammalian cell bias codon is optimized. Thoseaasigna virus 2A (T2A) self-cleavable peptide with high shearing efficiency is inserted between the TRE sequence and the recombinant adenovirus cLYZ sequence, and after synthesis, a pMD-19T vector containing a target gene fragment is obtained. A TRE gene, a T2A gene and a cLYZ gene are subcloned onto a shutter vector pDC315-MCS-EGFP; a recombinant shuttle vector and an adenovirus big skeleton pBHGlox(delta)E1, 3cre co-transfect an HEK293 cell by utilizing a LipoFiterTM liposomes transfection method, so that homologous recombination is realized, and the recombinant adenovirus Ad-TRE-T2A-cLYZ-EGFP is obtained.

Description

technical field [0001] The invention belongs to the field of genetic bioengineering, and relates to the prediction, design and synthesis of dominant B / T cell epitopes of Staphylococcus aureus (Staphylococcusaureus), Escherichia coli (Escherichia coli) and Streptococcus (Streptococcus), and cLYZ gene in T2A Peptide-mediated construction of a co-expression recombinant adenovirus method, specifically, a method of using bioinformatics to obtain the co-expression of the triple epitope gene (TRE) of the main pathogenic bacteria of cow mastitis and the grouper c-type lysozyme gene cLYZ The method and application of recombinant adenovirus vector construction. Background technique [0002] Cow mastitis is a serious disease that endangers the production of dairy cows and is mainly caused by the cross or co-infection of Staphylococcus aureus, Escherichia coli and Streptococcus, which has been plaguing the healthy development of the dairy industry in the world. At this stage, it mainly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N7/01A61K38/47A61P15/14A61P31/04A61K38/16
CPCA61K38/164A61K38/47C07K14/245C07K14/31C07K14/315C12N7/00C12N9/2462C12N15/86C12N2710/10021C12N2710/10043C12N2800/22C12Y302/01017A61K2300/00
Inventor 金天明林静李小慧高珂珂尚翠玲孟佳丽李昕赵瑞利
Owner TIANJIN AGRICULTURE COLLEGE
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