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Method for preparing plectasins by saccharomyces cerevisiae

A technology of mycelia and Saccharomyces cerevisiae, which is applied in the field of biotechnology and genetic engineering, and can solve the problems of residual methanol, harm, and high cost

Active Publication Date: 2014-12-24
CHANGSHA ZHONGKEJINGBO BIOTECH
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to overcome the deficiencies in the prior art, such as the difficulty of separation and purification, long production cycle, high cost, harm caused by residual methanol, and low product activity, the present invention provides a method for expressing the antimicrobial peptide Mycelia by using Saccharomyces cerevisiae prime method

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  • Method for preparing plectasins by saccharomyces cerevisiae
  • Method for preparing plectasins by saccharomyces cerevisiae
  • Method for preparing plectasins by saccharomyces cerevisiae

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Embodiment Construction

[0102] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments, without limiting the present invention.

[0103] like figure 1 Shown, the method for producing Plectasin with Saccharomyces cerevisiae of the present invention, it may further comprise the steps:

[0104] a. Acquisition of the promoter Ppgk gene, α-signal peptide gene and Plectasin gene: clone the promoter Ppgk gene, the Ppgk gene is from Nanyang K strain, with a full length of 793bp; clone the α-signal peptide gene, the α -The signal peptide gene is from the plasmid pPIC9K, with a full length of 277bp; the synthetically modified Plectasin gene, with a full length of 123bp;

[0105] b. Constructing the recombinant plasmid pYES2-CPS-Plec expressing plectasin gene in yeast cells;

[0106] c. Transforming Saccharomyces cerevisiae INVSc1 cells with the recombinant plasmid pYES2-CPS-Plec, the method of transforming the recombinant plasmid is elec...

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Abstract

The invention discloses a method for preparing plectasins by saccharomyces cerevisiae. The method comprises the following steps: a, transforming and amplifying saccharomyces cerevisiae genes; b, building recombinant plasmids pYES2-CPS-Plec of the saccharomyces cerevisiae genes in yeast cells; c, transforming INVSC1 cells of saccharomyces cerevisiae genes by the recombinant plasmids pYES2-CPS-Plec; d, screening positive bacteria strains, and performing digestion and identification; e, cultivating transformed bacteria to obtain antimicrobial peptide saccharomyces cerevisiae genes; and f, identifying the activity of the antimicrobial peptide saccharomyces cerevisiae genes. The plectasins provided by the invention has an obvious bacteriostatic action on a saccharomyces cerevisiae expression system, and the antibacterial effect is stronger than that of the antibacterial effect before modification. The saccharomyces cerevisiae expression system provided by the invention has the following advantages of being better in antibacterial effect, fast in growth rate and high in yield, is likely to realize high-density culture and does not need adopt methanol induction; furthermore, a protein product for expressing secretion is likely to be separated and purified; moreover, the saccharomyces cerevisiae expression system has a plurality of posttranslational modification features, such as polypeptide folding, glycosylation, methylation, acetylation and the like; in addition, due to a clear genetic background, the saccharomyces cerevisiae expression system is likely to operate an expression vector.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and genetic engineering, in particular to a method for producing plectasin by using Saccharomyces cerevisiae. Background technique [0002] China is a large animal husbandry country, with an annual production of more than 100 million tons of animal feed, and the loss caused by feed corruption during storage exceeds 1 billion yuan. At present, the commonly used measure to solve this problem is to add chemical preservatives to the feed, but the chemical preservatives left in the animal body not only affect the quality of livestock and poultry products, but also threaten people's health, such as causing gastroenteritis, etc. Diseases that affect liver and kidney functions, etc. On the other hand, due to the long-term large-scale addition of antibiotics in animal feed in my country, it seriously affects the micro-ecological balance of the animal digestive tract, resulting in the occurrence of ...

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Application Information

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IPC IPC(8): C12P21/02C12N15/81C12N15/66C12R1/865
Inventor 夏新界崔延春
Owner CHANGSHA ZHONGKEJINGBO BIOTECH
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