Isolation and identification for bombyx mori intestinal tract probiotics, and cloning and expression of enzyme gene produced by bombyx mori probiotics

A technology of intestinal probiotics and silkworm, which is applied in genetic engineering, plant genetic improvement, and microbial-based methods, etc., can solve the problems of few research reports on gut-derived α-amylase-producing bacteria, and achieve the improvement of the intestinal tract of silkworms The effect of environment and good application prospects

Inactive Publication Date: 2011-11-16
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

At present, there is no application of gut-derived Bacillus cereus and the enzymes it produces in sericulture m

Method used

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  • Isolation and identification for bombyx mori intestinal tract probiotics, and cloning and expression of enzyme gene produced by bombyx mori probiotics
  • Isolation and identification for bombyx mori intestinal tract probiotics, and cloning and expression of enzyme gene produced by bombyx mori probiotics
  • Isolation and identification for bombyx mori intestinal tract probiotics, and cloning and expression of enzyme gene produced by bombyx mori probiotics

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Embodiment Construction

[0030] 1. Selection of materials and preparation of medium

[0031] 11. Bombyx mori gut samples The silkworm Bombyx mori Dazao variety was provided by the Sericulture Department of Anhui Agricultural University, and the silkworms fed to the fifth instar by artificial diet were used as test materials.

[0032] 1.2 Test medium NA medium: peptone 10.0g, beef extract 5.0g, NaCl 5.0g, agar 18.0g, water 1L, pH 7.2-7.4. NA starch screening medium: NA medium supplemented with 1.0% soluble starch. LB medium: tryptone 10g, yeast extract 5g, NaCl 10g, water 1L, pH 7.0, solid medium added with 1.5% agar powder, autoclaved for later use. Liquid medium without agar.

[0033] 1.3 Main reagents and plasmid K1201 UNIQ-10 Column Bacterial Genomic DNA Extraction Kit, DNA molecular weight markers, PCR amplification reagents, etc. were purchased from Shanghai Sangon Bioengineering Co., Ltd.; dNTPs, competent cells Trans5α, pEASYT1, Transetta, etc. were purchased From TransGen; Plasmid Extractio...

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Abstract

The invention discloses a method for isolating and identifying bombyx mori intestinal tract probiotics, and cloning and expression of enzyme gene produced by bombyx mori intestinal tract probiotics. With the present invention, bacillus cereus in the bombyx mori intestinal tract is isolated and identified, the enzyme gene produced by the bacillus cereus is cloned and expressed; the method provides important significance for improving intestinal environment of the bombyx mori, and raising digestibilities of mulberry leaf and feed starch, and leaf-silk conversation rate. The bombyx mori intestinal tract probiotics capable of enzyme producing is screened through the method provided by the present invention, belongs to the range of feed-grade microbial feed additives, and has good application prospect.

Description

technical field [0001] The invention relates to the isolation and identification of silkworm intestinal probiotics and a method for cloning and expressing enzyme genes thereof, belonging to the field of biological technology. Background technique [0002] The animal gut is an ideal place for the growth and reproduction of a variety of microorganisms. Normal intestinal microbes have nutritional, immune, growth-stimulating, and biologically antagonistic effects on the host, and are a physiological component of the body. Silkworm is an important economic insect with huge economic and social benefits. However, there is always a low silk conversion rate (about 10%) in the sericulture production process, and the cost of mulberry leaves accounts for more than 50% of the silkworm cocoon production cost. Simultaneously, the incidence rate of silkworm disease is high, and the amount of cocoons lost due to silkworm disease in my country reaches more than 10% every year. Therefore, i...

Claims

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Application Information

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IPC IPC(8): C12N9/28C12N15/56C12N15/63C12N1/21C12N1/20C12R1/085C12R1/19
Inventor 王在贵刘朝良杨文静韩薇
Owner ANHUI AGRICULTURAL UNIVERSITY
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