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Antibiotic peptide gene and its yeast expression carrier

An expression vector and antibacterial peptide technology, which are applied in the field of research of fish antibacterial peptide gene and yeast recombinant expression vector

Inactive Publication Date: 2006-05-31
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

There is no report on the construction of fish antimicrobial peptide gene recombinant expression vector and its expression in yeast

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  • Antibiotic peptide gene and its yeast expression carrier

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Embodiment Construction

[0023] Taking the cloning of the turbot antimicrobial peptide hepcidin gene and the construction of an expression vector as an example, the technical content of the present invention will be described in detail in conjunction with the accompanying drawings.

[0024] The technical content of the present invention is as follows: 1. Cloning of turbot antimicrobial peptide (hepcidin) gene; including cDNA cloning, amplification of genomic DNA sequence, analysis and comparison of genomic DNA sequence. 2. Construction of recombinant hepcidin gene yeast expression vector and its expression in yeast: including construction of expression vector, transformation of expression vector into yeast cells, screening of recombinant yeast containing turbot antimicrobial peptide gene, recombinant antimicrobial peptide small Quantitative expression.

[0025] 1. Cloning of turbot antimicrobial peptide hepcidin gene:

[0026] 1. Amplification and cloning of cDNA: TRIzol kit was used to extract total...

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Abstract

A scophthalmus maximus peptide hepcidin gene and its yeast expression vector, the method is to separate and clone Scophthalmus maximus peptide cDNA and ití»s gene groupí»s gene, regroup this geneí»s mature peptides to yeast cell expression vector®ñpGAPZB, then to form confluent expression vector®ñpGAPZB-the6his, and conversion Pichia pastoris strain SMD1168ú¼and get the yeast plant that can express mature peptide albumen steadily, this peptide gene can be considered to finish expression by passing the SDS-PAGE and Western blot test. This invention clone Scophthalmus maximums peptide gene and form mature yeast expression vector in the cell at the first time, it can be used as fish and other aquatic animalí»s feed additive by yeast form directly, and it doní»t request to separate and purificate the regroup peptideíúThe invention also can be used to clone many fishí»s peptidegene and its expression vector, and used to prevention and cure fish and other aquatic animalí»s disease.

Description

Technical field: [0001] The invention belongs to the fish gene engineering technology in the field of biotechnology, and relates to a method for developing a fish antibacterial peptide gene and a yeast recombinant expression vector that can be used in fish disease prevention and control. Background technique: [0002] With the widespread use of antibiotics, the problem of drug resistance of pathogenic bacteria has attracted more and more attention, and the development of new antibacterial drugs is imminent. Fish is a vertebrate with both specific and nonspecific immunity. Nonspecific immunity plays an important role in the defense response of fish to external stimuli and pathogenic organisms. Antimicrobial peptides are a class of non-specific immune factors that can resist a variety of pathogenic infections. Studies have found that antimicrobial peptides play an important role in the process of fish and shrimp resisting pathogenic infection. There are many antimicrobial p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12N15/10C12N15/81C12P21/02
Inventor 陈松林李伟
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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