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Fish-derived protease gene and application thereof

A protease and fish-derived technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of large-scale cultivation of bacteriophages and viruses, achieve promising prospects, promote digestion and absorption, increase weight gain rate and specific growth rate effect

Active Publication Date: 2021-12-10
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many potential dangers in displaying proteins on the surface of conditional pathogenic bacteria and pathogenic bacteria. In addition, it is difficult to cultivate phages and viruses on a large scale

Method used

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  • Fish-derived protease gene and application thereof
  • Fish-derived protease gene and application thereof
  • Fish-derived protease gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Screening of fish-derived protease-producing strains, screening of protease-producing genes and construction of recombinant plasmids

[0046] This embodiment includes: screening protease-producing strains from the intestinal tract of California perch, then analyzing and screening protease-producing genes from the strains, and constructing recombinant plasmids comprising the screened protease-producing genes, specifically including the following steps:

[0047] (1), screening of protease-producing strains

[0048] Screening of solid medium (recipe: peptone 1g / 100mL, yeast extract 0.5g / mL, sodium chloride 1g / mL, technical agar powder 1.5g / 100mL, skimmed milk powder 1g / 100mL, pH7.2) by protease screening The intestinal strain of California perch was screened to observe whether there was a transparent enzyme-producing circle around the colony. If there is a transparent enzyme-producing circle to appear, it indicates that it is a protease-producing bacterial strai...

Embodiment 2

[0100] Example 2 Effects of Three Yeast Recombinant Expression Vectors Containing Recombinant Plasmids on the Growth Performance of Farmed Fish

[0101] This embodiment uses the three yeast recombinant expression vectors containing recombinant plasmids of Example 1 to test, specifically including the following steps:

[0102] 1. Test groups

[0103] A large blue tank containing 400L water (average water temperature 26°C-28°C) was used to carry out the California perch animal experiment. Three experimental groups and one control group were set up, with 90 fish in each group, and a total of 3 parallel fish were set up, that is, 30 fish in each parallel. Fish, with control group 1, control group 2, control group 3, experiment group 1 (ORF32-1, ORF32-2, ORF32-3), experiment group 2 (ORF436-1, ORF436-2, ORF436 respectively) in the following examples -3) and experimental group 3 (ORF160-1, ORF160-2, ORF160-3).

[0104] 2. Yeast count and bacterial quantity setting

[0105] After ...

Embodiment 3

[0127] Example 3 Effects of Three Yeast Recombinant Expression Vectors Containing Recombinant Plasmids on Fish Protease Activity

[0128] The grouping and feeding methods of Example 2 were used to culture California perch. After 28 days, the intestinal tissues of 3 fish were taken in parallel in each group, and the mixed sample homogenate was diluted 5 times, and then the enzyme activity was measured. The pepsin activity of each group of fish was measured by Solarbio pepsin activity detection kit (article number: BC2325 specification: 100T / 48S), and the activity of each group of fish was measured by Solarbio trypsin activity detection kit (article number: BC2315 specification: 100T / 96S). Trypsin activity. For the enzyme activity determination operation steps and the enzyme activity calculation formula, refer to the kit instructions.

[0129] 1. After 28 days of breeding, detection of pepsin activity in the intestinal tract of California sea bass (pepsin kit)

[0130] Enzyme ...

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Abstract

The invention discloses a fish-derived protease gene and application thereof. An open reading frame of the fish-derived protease gene has a nucleotide sequence shown as SEQ ID No.1, SEQ ID No.3 or SEQ ID No.5, or is a nucleotide sequence complementarily paired with SEQ ID No.1, SEQ ID No.3 or SEQ ID No.5, or is a nucleotide sequence with a coding amino acid sequence shown as SEQ ID No.2, SEQ ID No.4 or SEQ ID No. 6. The fish-derived protease gene disclosed by the invention is beneficial to promoting the digestive absorption capacity of fish on high-protein artificial feed, improves the growth performance of the fish, can be specifically applied to fish culture, and is green, efficient and considerable in prospect.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and more specifically, the invention relates to a fish-derived protease gene, a recombinant plasmid comprising the fish-derived protease gene, a yeast recombinant expression vector comprising the recombinant plasmid, and its expression in fish Application in farming and improving fish digestibility. Background technique [0002] The gut of fish is an important site for food digestion and nutrient absorption. After fish ingest, the nutrients of the food depend on the action of various enzymes. Proteins and polypeptides in food need to be hydrolyzed by proteases in digestive juice or intestinal wall cells to become amino acids that intestinal cells can absorb. Therefore, the activity of intestinal protease plays an important role in the growth and metabolism of fish. [0003] The current status of fish farming is: under the continuous feeding of high-protein artificial feed, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/52C12N15/57C12N15/81A23K50/80A23K10/16
CPCC12N9/52C12N15/81A23K50/80A23K10/16Y02A40/81
Inventor 冯国清刘振兴梁志凌马江耀郝乐马艳平
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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