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Construction method of recombinant adenovirus co-expressing sal-2 and hlyz dual antibacterial genes, recombinant adenovirus and application

A technology of recombinant adenovirus and gene recombination, applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of difficult treatment of bacterial drug-resistant regional cow mastitis

Active Publication Date: 2018-03-09
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

At present, the prevention and treatment of the disease is mainly through standardized breeding management and the use of antibiotics and chemotherapy drugs. Difficulties in the treatment of regional cow mastitis

Method used

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  • Construction method of recombinant adenovirus co-expressing sal-2 and hlyz dual antibacterial genes, recombinant adenovirus and application
  • Construction method of recombinant adenovirus co-expressing sal-2 and hlyz dual antibacterial genes, recombinant adenovirus and application
  • Construction method of recombinant adenovirus co-expressing sal-2 and hlyz dual antibacterial genes, recombinant adenovirus and application

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Embodiment Construction

[0023] Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail:

[0024] With the development of molecular biology, immunology, pathology and related disciplines, gene therapy has become a hot spot in the research of disease prevention and treatment. At present, adenovirus (adenovirus, Ad) vector therapy is one of the most promising gene transfer methods in gene therapy. It can effectively deliver foreign genes to various target cells or tissues. The construction and operation of the vector are simple, It has a wide range of hosts and strong infectivity. It can enter different tissues through different channels, and can also infect differentiated non-dividing cells.

[0025] A large number of research results show that endolysin (virolysin) is the main component of bacteriophage to achieve the lysis function after infecting the host bacteria. It achieves the purpose of lysing bacteria by hydrolyzing the connec...

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Abstract

The invention discloses a construction method of a recombinant adenovirus vector co-expressing SAL‑2 gene and hLYZ dual antibacterial gene, and the application of the recombinant adenovirus. The extra SAL‑2 gene and hLYZ gene were cloned into the pShuttle‑IRES‑hrGFP‑2 shuttle vector, linearized and recombined with the shuttle vector and co-transfected with the backbone vector pAdeasy‑1 into E. coli BJ5183 competent cells , making it perform homologous recombination in BJ5183, constructing the recombinant adenoviral plasmid Ad-SAL-LYZ containing the above-mentioned target gene; digesting the recombinant adenoviral plasmid with PacⅠ, and transfecting HEK293 cells through liposome-mediated transfection, High-titer recombinant adenovirus was packaged to obtain high-titer recombinant adenovirus; the joint inhibitory effect of SAL‑2 gene and hLYZ gene on the main pathogenic bacteria of dairy cow mastitis was evaluated under the mediation of adenovirus.

Description

technical field [0001] The invention relates to a method for constructing a recombinant adenovirus, specifically, a method for constructing a recombinant adenovirus co-expressing SAL-2 and hLYZ dual antibacterial genes, and the recombinant adenovirus and its application. Background technique [0002] Cow mastitis, also known as cow mastitis, is an inflammatory reaction that occurs when the cow's mammary gland is stimulated by the outside world or infected by pathogenic bacteria. It will not only cause a decline in milk production of dairy cows, shorten the replacement cycle of dairy cows, and even cause the complete loss of lactation ability of sick cows in severe cases. Therefore, the clinical prevention and treatment of mastitis in dairy cows has been a major problem that has plagued the dairy farming industry. At present, the prevention and treatment of the disease is mainly through standardized breeding management and the use of antibiotics and chemotherapy drugs. It b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/861C12N15/66C12N7/01A61K35/761A61P31/04
Inventor 金天明李萌龙雨清张东超弓建芳高珂珂李小慧林静
Owner TIANJIN AGRICULTURE COLLEGE
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