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Cascade expression of active fragment of cholecystokinin and application thereof

A technology of cholecystokinin and active fragments, applied in the field of genetic engineering

Inactive Publication Date: 2007-08-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report of expressing CCK or its active fragments (such as CCK-33 peptide, CCK-8 peptide) in Escherichia coli

Method used

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  • Cascade expression of active fragment of cholecystokinin and application thereof
  • Cascade expression of active fragment of cholecystokinin and application thereof
  • Cascade expression of active fragment of cholecystokinin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0060] 1. Construction of recombinant plasmid expressing cholecystokinin 33 peptide

[0061] Firstly, according to the base sequence of the chicken cck gene and high-frequency codons of Escherichia coli, the cck-33 gene was designed and synthesized. Then, insert the cck-33 gene into the pRSETA plasmid to obtain the recombinant plasmid pRSETA-1CCK, the specific operation is as follows:

[0062] Specific primers P1 (5'-CATGGATCCGGTTCTACTGGCCGCT-3') / P2 (5'-CATGAGCTCCCAAAATCCATCCAGCC-3') were designed according to the artificially synthesized cck-33 gene, and the PCR method was used to amplify the artificially synthesized cck-33 gene as a template. Increase cck-33 gene fragment. The PCR reaction system (200 μL) for amplifying the cck-33 gene fragment was composed as follows: 20 μL 10×buffer, 16 μL dNTPs, 2 μL Taq enzyme, 1.5 μL primer P1, 1.5 μL primer P2, 0.5 μL artificially synthesized cck-33 gene template, and finally Add water to a total volume of 200 μL. The PCR temperatur...

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Abstract

This invention provides a method for series expression of active fragment of cholecystokinin, it concludes recombined plasmid construction of expression cholecystokinin gene, expression in large intestine bacilli of cholecystokinin gene and pure proceed of outcome in the project of cholecystokinin gene. Plasmid of expression cholecystokinin gene has chick cholecystokinin 33 gene or many series sequence of nucleotide. The outcome of the project of cholecystokinin gene has chick cholecystokinin 33 peptide or many series sequence of residue of amino acid.

Description

technical field [0001] The invention belongs to the field of genetic engineering. It specifically relates to the construction of a recombinant plasmid expressing cholecystokinin gene, the expression of cholecystokinin gene in Escherichia coli and the purification of cholecystokinin gene engineering product. The present invention further relates to the active immunization technology and application of the recombinant cholecystokinin polypeptide to pigs, rabbits, cows, chickens and other animals. Background technique [0002] Cholecystokinin (CCK) is an effective factor regulating animal feed intake. Cholecystokinin, also known as cholecystokinin or trypsin, is a polypeptide hormone secreted by I cells of the small intestinal mucosa. The precursor of CCK is 130 amino acids, which are gradually decomposed into small molecules of CCK. The CCKs discovered so far include CCK-83, CCK-58, CCK-39, CCK-33, CCK-12, CCK-8 and CCK-4. Its distinct sites are mainly at the carboxy-termin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/69C07K14/465A61K39/00C07K16/18G01N33/53A23K1/00A23K1/16A61P1/00A23K20/147A23K50/10A23K50/30A23K50/50A23K50/75
Inventor 曹永长覃健萍毕英佐舒鼎铭马静云谢青梅
Owner SOUTH CHINA AGRI UNIV
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