Synthesized duck beta-defensins-2 gene, recombinant plasmid containing gene as well as production methods for recombinant yeast transformant and recombinant duck beta-defensins-2 protein

Abstract

The invention discloses a synthesized duck beta-defensins-2 gene, a recombinant plasmid containing the gene as well as production methods for a recombinant yeast transformant and a recombinant duck beta-defensins-2 protein. The recombinant duck beta-defensins-2 gene refers to the cDNA (complementary Deoxyribonucleic Acid) gene sequence of a duck AvBD2 gene registered in GenBank; meanwhile, the nucleotide sequence in a mature peptide segment of the duck AvBD2 gene is transformed according to the preference of a yeast codon; the duck beta-defensins-2 gene is synthesized by using a gene synthetic method; the construction process of the recombinant plasmid containing the gene sequentially comprises the following steps of: firstly, setting an upstream primer and a downstream primer of the duckbeta-defensins-2 gene; secondly, carrying out PCR (Polymerase Chain Reaction) amplification on a duck beta-defensins-2 gene segment; and thirdly, constructing a recombinant plasmid pPICZalpha-A-AvBD2. The production method for the recombinant duck beta-defensins-2 protein comprises the following steps of: firstly, preparing and linearizing the recombinant plasmid; secondly, preparing a competent cell obtained by electrotransformation of pichia pastoris; and thirdly, electroporating the pichia pastoris. The production method for the recombinant duck beta-defensins-2 protein comprises the step of induced expression of the positive yeast transformant. Compared with the prior art, synthesized duck beta-defensins-2 gene, the recombinant plasmid and the production methods disclosed by the invention have the advantages of activities of resisting bacteria and promoting the growth, suitability for application to the livestock production, low production cost and high production efficiency.

Description

technical field [0001] The invention relates to a gene synthesis, recombinant plasmid construction, recombinant yeast transformant acquisition and recombinant protein 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 defen...

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