Construction and application of artificially-synthesized chicken interferon gene sequence and recombinant engineering bacterium thereof
A technology of recombinant engineering bacteria and chicken interferon, which is applied in the field of biological engineering, can solve the problems of large yeast fermentation output and drug residues, and achieve the effects of easy fermentation, large output, and improving the body's immunity
Inactive Publication Date: 2012-12-05
GUIZHOU UNIV
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[0004] Aiming at the defects in the above-mentioned fields, the present invention provides an artificially synthesized chicken interferon gene sequence, and provides recombinant engineering bacteria obtained by using Saccharomyces cerevisiae as a recipient bacterium, and using Saccharomyces cerevisiae as a recipient bacterium to express chicken interferon intracellularly , the yield of yeast fermentation is large, the product does not need to be separated and purified, and the recombinant strain of yeast feed containing chicken interferon can be directly fed to the recombinant strain, which can realize the low-cost supply of interferon in animal husbandry, and improve the immunity of chicken while providing nutrients To avoid the cumbersome management of vaccination and other drug residues in livestock products caused by feeding antibiotics and other drugs
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[0047] The present invention will be described in further detail below in conjunction with embodiment.
[0048] SC selection medium is: 0.8% Ura deficient medium powder, 2% glucose
[0049] YPD screening medium is: 2% tryptone, 1% yeast extract, 2% glucose (solid medium plus 2% agar powder), 60mg / l Amp.
[0050] Induction medium: 0.8% Ura deficient medium powder, 2% galactose, 1% raffinose;
[0051] Or 0.8% Ura deficient medium powder, 2% galactose, 2% glucose
[0052] (1) Materials and methods
[0053] 1. Plasmids and strains
[0054] Saccharomyces cerevisiae expression plasmid pYES2 / CT and Saccharomyces cerevisiae strain INVSc1 were purchased from Invitrogen; Escherichia coli strain DH5α was preserved by the Key Laboratory of Agricultural Bioengineering of Guizhou Province. It is a common strain and can be distributed to the public.
[0055] 2. Saccharomyces cerevisiae expression vector construction
[0056] According to the sequences of the interferon genes ChIFN-α (Ge...
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Abstract
The invention relates to construction and application of an artificially-synthesized chicken interferon gene sequence and a recombinant engineering bacterium thereof, and belongs to the field of biological technology. According to the invention, an artificially-synthesized sequence such as SEQ ID NO1 or SEQ ID NO2 is adopted, and saccharomyces cerevisiae is used as a host bacterial strain to obtain the recombinant engineering bacterium; detection results from PCR, restriction enzyme digestion, sequencing, SDS-PAGE, ELISA and CPE show that: ChIFN-alpha and ChIFN-gamma genes can be expressed inthe host bacterial strain, and the expressed protein has significant biological activity. Experiment shows that by using saccharomyces cerevisiae as a bioreactor to produce chicken interferon pharmaceutical protein, not only the production cost of interferon is greatly decreased, but also high-quality protein is provided; at the same time, the purpose of body immune function improvement is reached; and the invention lays a foundation for animal epidemic disease prevention and treatment by using yeast additives.
Description
technical field [0001] The invention relates to the field of biological processes, in particular to an artificially synthesized chicken interferon gene sequence, a recombinant engineered bacterium obtained by using Saccharomyces cerevisiae as a recipient bacterium, which can express intracellularly and produce animal interferon. Background technique [0002] Saccharomyces cerevisiae is a good nutritional and health-care feed additive. It is rich in nutrition, rich in B vitamins, minerals, digestive enzymes, growth-promoting factors and relatively complete amino acids. The use of yeast additives can increase the protein content of the feed, stimulate the growth of beneficial bacteria, improve the body's immunity, promote the digestion and absorption of nutrients and the growth and development of animals. At the same time, Saccharomyces cerevisiae is also the earliest yeast strain used in genetic engineering, and it is a model organism for the study of eukaryotic biology. The ...
Claims
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IPC IPC(8): C12N15/21C12N15/23C12N15/81C12N1/19C12P21/02C12R1/865
Inventor 赵德刚宋莉卢凌霄
Owner GUIZHOU UNIV