Recombinant adenovirus capable of co-expressing double antibacterial genes of SAL-2 and hLYZ, and construction method and application thereof

A technology of SAL-2 and recombinant adenovirus, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems such as the difficulty in the treatment of bacterial drug-resistant regional cow mastitis

Active Publication Date: 2015-12-09
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

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  • Recombinant adenovirus capable of co-expressing double antibacterial genes of SAL-2 and hLYZ, and construction method and application thereof
  • Recombinant adenovirus capable of co-expressing double antibacterial genes of SAL-2 and hLYZ, and construction method and application thereof
  • Recombinant adenovirus capable of co-expressing double antibacterial genes of SAL-2 and hLYZ, and construction method and application thereof

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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 recombinant adenovirus capable of co-expressing double antibacterial genes of SAL-2 and hLYZ, and a construction method and application thereof. The construction method includes amplifying the SAL-2 gene and the hLYZ gene from a staphylococcus aureus phage genome and a human milk somatic cell total RNA (ribonucleic acid) respectively, cloning the obtained target genes into a shuttle vector pShuttle-IRES-hrGFP-2, co-transfecting an escherichia coli BJ5183 competent cell by the shuttle vector and a framework vector pAdeasy-1 after linear recombination of the shuttle vector to achieve homologous recombination in BJ5183, and constructing a recombinant adenovirus plasmid AD-SAL-LYZ containing the target genes; transfecting an HEK293 cell through liposome mediation after Pac I digestion of the recombinant adenovirus plasmid, and packing to obtain the high-titer recombinant adenovirus; evaluating the joint inhibiting effect of the SAL-2 gene and the hLYZ gene on main pathogenic bacteria of cow mastitis under adenovirus mediation.

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 Applications(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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