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Ssc-miR-122-5p related to piglet C-type clostridium perfringens infectious diarrhea, and application of ssc-miR-122-5p

A technology for Clostridium perfringens and infectious diarrhea, applied in ssc-miR-122-5p and its application fields, can solve the problem of less research on the mechanism of action

Active Publication Date: 2021-09-28
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there are few studies on the mechanism of action of miRNA in bacterial infectious diarrhea in piglets.

Method used

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  • Ssc-miR-122-5p related to piglet C-type clostridium perfringens infectious diarrhea, and application of ssc-miR-122-5p
  • Ssc-miR-122-5p related to piglet C-type clostridium perfringens infectious diarrhea, and application of ssc-miR-122-5p
  • Ssc-miR-122-5p related to piglet C-type clostridium perfringens infectious diarrhea, and application of ssc-miR-122-5p

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1, expression detection of ssc-miR-122-5p

[0021] 1. Expression detection of ssc-miR-122-5p in ileal tissue of piglets with Clostridium perfringens type C infectious diarrhea

[0022] Step 1. Sample collection

[0023] Select 30 healthy 7-day-old piglets (long × large), randomly select 5 piglets as the control piglet group (IC), and feed the same dose of culture solution that was not inoculated with the strain, and the remaining 25 piglets were treated with Clostridium perfringens type C Bacteria oral challenge test, each piglet was fed with culture solution of Clostridium perfringens type C C59-2 strain (1×10 9 CFU / mL) 1 mL, the test period was 5 days. The ileal tissue samples were quickly collected and stored in liquid nitrogen.

[0024] Step 2. Extraction of total RNA from ileum tissue

[0025] Take out the tissue sample from liquid nitrogen, grind it into powder, collect it in a 1.5 mL centrifuge tube, add 1 mL TransZol Up, shake and mix well, let stand...

Embodiment 2

[0045] Example 2, Effects of overexpression and interference of ssc-miR-122-5p on inflammatory injury of IPEC-J2 cells treated with CPB2 toxin

[0046] 1. Overexpression of ssc-miR-122-5p and detection of interference efficiency

[0047] Step 1. IPEC-J2 cell culture

[0048] Step is with embodiment 1.

[0049] Step 2. Chemical synthesis of miRNA mimics and inhibitors

[0050] According to the sequence information of porcine ssc-miR-122-5p provided by miRBase database, miRNA mimics and inhibitors were chemically synthesized in vitro. The synthesized small RNA fragments were designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. The synthetic sequence of ssc-miR-122-5pmimic is shown in SEQ ID NO.3-SEQ ID NO.4, and the synthetic sequence of ssc-miR-122-5p inhibitor is shown in SEQ ID NO.5.

[0051] Step 3, cell transfection

[0052] IPEC-J2 cells were seeded in 6-well culture plates, and when the cell density reached 70%, they were washed 2-3 times wi...

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Abstract

The invention discloses ssc-miR-122-5p related to piglet C-type clostridium perfringens infectious diarrhea, and the nucleotide sequence of the ssc-miR-122-5p is as shown in SEQ ID NO. 1. The invention also discloses a specific primer for detecting the expression quantity of the ssc-miR-122-5p, and the sequence of the specific primer is as shown in SEQ ID NO. 2. The invention discloses a method for improving the C type clostridium perfringens infection resistance of piglets by interfering ssc-miR-122-5p. The invention discovers the obvious high expression of ssc-miR-122-5p in the ileum tissues of the piglet C-type clostridium perfringens infectious diarrhea for the first time, then, IPEC-J2 cells are taken as a research object, and the influence of the ssc-miR-122-5p on IPEC-J2 cell injuries induced by CPB2 toxins is researched through overexpression and interference. The invention aims to clarify the action mechanism of ssc-miR-122-5p in the process of regulating and controlling diarrhea caused by C-type clostridium perfringens infection, and provides a powerful scientific research reference for molecular breeding of bacterial diarrhea of piglets.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to ssc-miR-122-5p related to piglet C-type Clostridium perfringens infectious diarrhea and application thereof. Background technique [0002] At present, Clostridium perfringens type C has become the main cause of intestinal infection of various livestock and poultry in modern farming industry, especially for piglets under one week old, it can cause hemorrhagic diarrhea, and then the bacteria and toxins enter the whole body circulatory system, leading to multi-organ failure and eventually death of the organism. According to statistics, the morbidity and mortality of piglet diarrhea caused by Clostridium perfringens type C can be as high as 100%. At present, the incidence of piglet diarrhea has been reduced by means of antibiotics and vaccines, and with the excessive use of antibiotics, not only the structure of the intestinal mucosa of piglets has been changed, the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113C12N15/11A61K31/7088A61P1/12A61P31/04
CPCC12Q1/6883A61K31/7088A61P1/12A61P31/04C12Q2600/158C12Q2600/178
Inventor 张生伟滚双宝高小莉杨巧丽
Owner GANSU AGRI UNIV
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