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Production method of recombinant yeast positive transformant of synthesized recombinant duck beta-defensin-2 protein recombinant yeast positive transformant

A technology of recombinant yeast and defensins, which is applied in the field of animal biomedical engineering, can solve the problems of complicated procedures, high cost, and limitations, and achieve the effects of increasing expression, facilitating maintenance, and low production costs

Active Publication Date: 2013-07-17
KAIPING CITY LVHUANG AGRI & STOCKBREEDING DEV
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0003] Due to the low content of natural defensins in animals, direct extraction is far from meeting the needs, and the cost is high, the yield is low, and the process is complicated; chemical synthesis is feasible but the cost is too high, and the application is limited; with the development of genetic engineering technology, through genetic recombination It is possible to produce defensin peptides with technology to meet the needs of research and production

Method used

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Embodiment Construction

[0027] Now in conjunction with embodiment the present invention is described in further detail:

[0028] The duck β-defensin-2 (AvBD2) gene nucleotide sequence design and in vitro synthesis process of the present invention are realized in this way,

[0029] Referring to the cDNA gene sequence of the duck AvBD2 gene registered in GenBank, the nucleotide sequence of the mature peptide of the duck AvBD2 gene was modified according to the yeast codon preference, and then synthesized by gene synthesis (sent to Shanghai Sangong Bioengineering Technology Co., Ltd. gene synthesis), the nucleotide sequence of the duck β-defensin-2 (AvBD2) gene synthesized after transformation is as follows:

[0030] GATATGTTTTTGTGTAGAATTGGTTCTTGTCATTTTGGTAGATGTCCAATTCATTTGGTTAGAGTTGGTTCCTGTTTTGGTTTTAGATCTTGTTGTAAGTCTCCATGGGATGTTTAA.

[0031] The recombinant plasmid pPICZα-A-AvBD2 containing the gene of the present invention is realized in this way,

[0032] The construction process of the recombinant...

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Abstract

A method for producing a recombinant yeast positive transformant of a synthesized recombinant duck beta-defensin-2 protein. The method is characterized by comprising the steps of: (1) preparation and linearization of a recombinant plasmid; (2) preparation of Pichia pastoris electroporation competent cells; and (3) electric shock transform of the Pichia pastoris: construction of a recombinant plasmid pPICZalpha-A-AvBD2. Compared with the prior art, the positive transformant provided by the invention has antibacterial and growth promoting activity, is suitable for application to animal production, and has the advantages of low production cost and high production efficiency.

Description

technical field [0001] The invention relates to a gene production method in the field of animal biomedical engineering. Background technique [0002] Animals used in modern animal husbandry are increasingly sensitive to environmental stress and pathogens while having high productivity. Especially under intensive feeding conditions, feeding animals face more stress and invasion of pathogens. There is an urgent need for a high-efficiency, non-toxic, non-residue immune enhancer and growth enhancer in animal husbandry production. While improving the disease resistance of feed animals, it can also improve the production performance of feed animals and the safety of animal products, so as to Promote the healthy development of animal husbandry and reduce the damage of animal husbandry to human health and the ecological environment. Previous studies have found that defensins have broad-spectrum bactericidal activity and super antimicrobial effect, and the mechanism of action is sp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N1/19C12R1/84
Inventor 张辉华韩小凤马保华毕英佐马静云谢青梅李辰
Owner KAIPING CITY LVHUANG AGRI & STOCKBREEDING DEV
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