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Preparation method of pig alpha-interferon compound preparation

The technology of a compound preparation and interferon, which is applied in the field of preparation of porcine alpha-interferon compound preparations, can solve the problems of difficult administration of the whole group, inability to administer oral administration, etc., and achieves low production cost and important clinical application value. , the effect of broad market prospects

Inactive Publication Date: 2013-09-04
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, as a glycoprotein cytokine, porcine IFN-α is easily digested by pepsin, so it can only be administered by intramuscular injection and cannot be administered orally. Difficulties with group dosing

Method used

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  • Preparation method of pig alpha-interferon compound preparation
  • Preparation method of pig alpha-interferon compound preparation
  • Preparation method of pig alpha-interferon compound preparation

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Embodiment Construction

[0051] A preparation method of porcine α-interferon compound preparation, its steps are as follows:

[0052] (1) Artificially synthesized 150bp short Lactobacillus signal peptide sequence, including the enzyme cleavage site upstream of SP N deI and downstream enzyme cutting sites, the downstream enzyme cutting sites are K pnI, S maI, Bam HI, X baI, S alI, Pst I. Sph I. Hin d III and Eco R I, and cloned into the PUCK vector to construct the pUCK-SP vector;

[0053] (2) A 501bp porcine IFN-α gene sequence was amplified from the pMD18-IFN-α vector by PCR, and inserted into the pUCK-SP vector K pnI and Eco R I restriction site, construct the pUCK-SP-IFN-α plasmid, pUCK-SP-IFN-α plasmid by N deI and Eco Digest with R I to recover the SP-IFN-α fragment;

[0054] (3) According to the Lactobacillus casei lactose operon gene sequence published by Genbank, design a pair of specific primers, and add the 5' end of the upstream primer P stI enzyme cutting site, the 5' ...

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Abstract

The invention discloses a preparation method of a pig alpha-interferon compound preparation. The preparation method comprises the steps as follows: (1) artificially synthesizing a small and short lactic acid bacillus signal peptide sequence of 150 bp and creating a pUCK-SP carrier; (2) creating pUCK-SP-IFN-alpha plasmids, carrying out enzyme digestion on the pUCK-SP-IFN-alpha plasmids through NdeI and EcoRI, and recovering an SP-IFN-alpha fragment; (3) creating lactobacillus casei replicate type expression plasmids pIAbeta8-plac-SP-IFN-alpha; and (4) obtaining the pig alpha-interferon compound preparation through an electric conversion method. According to the preparation method, a pig IFN-alpha gene is planned to be guided into lactobacillus casei so as to obtain lactobacillus casei capable of expressing pig IFN-alpha. A bacterial strain integrates the effects of virus resistance and bacteria resistance and can be very conveniently fed to animals in an oral administration way.

Description

technical field [0001] The invention relates to a preparation method of a porcine α-interferon compound preparation. Background technique [0002] my country is the country with the largest number of pigs in the world, and also the country with the most and most complicated pig diseases. At present, the poor control of important infectious diseases of pigs is still the main problem faced by the pig industry. Among the important infectious diseases of pigs, the economic losses caused by viral diseases, especially porcine reproductive and respiratory syndrome, porcine pseudorabies, porcine parvovirus disease, swine fever, swine flu, etc., are immeasurable. For these common viral diseases, although vaccines can be used for routine immunization, due to various reasons such as immunization procedures, vaccines themselves, and individual animals, the effect in actual production is not satisfactory, and the application of attenuated vaccines has the potential of returning to the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N1/21C12R1/245
Inventor 陈红英崔保安李坤马世杰魏战勇钞安军李新生
Owner HENAN AGRICULTURAL UNIVERSITY
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