Cytosine-guanine (CG) island oligonucleotide with immunostimulation capability for piglets and application of CG island oligonucleotide to feed additive

An oligonucleotide, immune stimulation technology, applied in the application, animal feed, animal feed, etc., can solve the problems of rapid adverse reactions, uncooperative piglets, tissue damage, etc., to improve the survival rate and breeding efficiency, reduce The effect of low morbidity and production cost

Inactive Publication Date: 2020-12-15
HANGZHOU HUADI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In addition, the current use of CG island oligonucleotides is usually absorbed as an immune adjuvant through body injection. During the injection process, piglets are prone to incompatibilities, and there are tissue damage, pain, potential complications, Adverse reactions occur rapidly and other problems

Method used

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  • Cytosine-guanine (CG) island oligonucleotide with immunostimulation capability for piglets and application of CG island oligonucleotide to feed additive
  • Cytosine-guanine (CG) island oligonucleotide with immunostimulation capability for piglets and application of CG island oligonucleotide to feed additive
  • Cytosine-guanine (CG) island oligonucleotide with immunostimulation capability for piglets and application of CG island oligonucleotide to feed additive

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 piglet rearing

[0031] Healthy piglets were selected. All piglets were purchased from the Institute of Biochemistry, School of Life Sciences and School of Medicine, Zhejiang Sci-Tech University, weighing 20-40kg, and all were raised in the breeding room of Building 6, Zhejiang Sci-Tech University. They were fed 3 times a day, and the feed was selected from the Logistics Support Co., Ltd. of Zhejiang Sci-Tech University. The breeding room is ventilated daily to ensure healthy breathing of piglets.

Embodiment 2

[0032] Embodiment 2 piglet infection model construction

[0033] A model of bacterial infection in piglets was constructed using porcine enterotoxic Escherichia coli. The piglets were divided into two groups, one was the disease group and the other was the normal group. Each piglet in the disease group was subcutaneously injected with 1 mL of porcine enterotoxic E. coli at a concentration of 10E5 / mL, and the normal group was used as a control by injecting the same dose of PBS.

Embodiment 3

[0034] Example 3 Preparation of pUC18-CG island oligonucleotide bacillus bacteria powder

[0035] The CG island oligonucleotide (SEQ ID NO.1) with immunostimulatory ability and the pUC18 empty vector (Hongye Xinchuang Antibody Technology Co., Ltd.) were ligated at a mass ratio of 1:3 by T4 ligase overnight at 16°C. The next day, the ligation product was transformed into a competent form of methylotrophic Bacillus (CMCC (B) 13337), and cultured on the Amp plate. After 12 hours, the plaques with obvious single colonies were selected and identified by sequencing, and the identified qualified pUC18-CG island oligonucleotide Bacillus strains were put into a fermenter for amplification and culture, and the fermentation product liquid was collected and freeze-dried to make bacterial powder. When used, it is mixed into pig feed at a ratio of 1:1000 (weight ratio).

[0036] The identified qualified pUC18-CG island oligonucleotide Bacillus strain was simultaneously sent to the General ...

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Abstract

The invention discloses cytosine-guanine (CG) island oligonucleotide with immunostimulation capability for piglets, and a sequence of the CG island oligonucleotide is as shown in SEQ ID NO.1. The invention further discloses a strain for expressing a CG island oligonucleotide vector; the strain is collected in China General Microbiological Culture Collection Center (CGMCC), and the collection number is 13337. Bacillus expressing the CG island oligonucleotide is made into dry powder; the dry powder is added into feed in the form of an additive, and the functions and effects of the dry powder aretested in animal bodies, so that a new solution is provided for disease treatment and prevention and cure of piglets.

Description

technical field [0001] The invention belongs to the technical development field of nucleic acid anti-infection preparations, in particular to the preparation of CG island oligonucleotides and their application in the field of piglet feed additives. [0002] technical background [0003] With the rapid development of my country's breeding industry, piglet diseases occur frequently and become more and more serious, causing huge economic losses. At the same time, due to the extensive use of antibiotics and insecticides, problems such as environmental pollution, pathogenic drug resistance and food safety have been caused, which have seriously affected the sustainable development of the aquaculture industry and people's health. With the country promulgating a series of strict food safety standards, a large number of antibiotics have been banned from being used in piglet breeding, and the breeding industry is facing a difficult situation where there is no medicine to do. Therefore...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/117C12N15/75C12N1/21A23K10/18A23K50/30A23K50/60C12R1/07
CPCA23K10/18A23K50/30A23K50/60C12N15/117C12N15/75C12N2310/17
Inventor 姜啸风胡智奕吕正兵周天琼周正兵何缤霞
Owner HANGZHOU HUADI GRP CO LTD
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