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A lncRNA ALDB-898 associated with piglet bacterial diarrhea and its application

A bacterial and diarrheal technology, applied in the fields of genetic engineering and pig disease resistance genetics, can solve the problem of less mechanism of action and achieve the effect of wide application value

Active Publication Date: 2022-07-29
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the mechanism of action of lncRNA in piglet diarrhea caused by Clostridium perfringens type C infection

Method used

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  • A lncRNA ALDB-898 associated with piglet bacterial diarrhea and its application
  • A lncRNA ALDB-898 associated with piglet bacterial diarrhea and its application
  • A lncRNA ALDB-898 associated with piglet bacterial diarrhea and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Expression and subcellular localization of lncRNA ALDB-898

[0034] 1. Expression of LncRNA ALDB-898

[0035] 1. LncRNA transcriptome sequencing

[0036]In this study, 30 7-day-old piglets (long × big) were selected, and 5 piglets were randomly selected as the control group (IC), fed with the same dose of culture medium without the inoculated strain, and the remaining 25 piglets were treated with C-type perfringens. Bacteria oral challenge test, each piglet was given 1 mL of C. perfringens C59-2 strain culture solution, and the test period was 5d. After the test, the sum of the defecation scores of each piglet during the test period was used as the total diarrhea score value of the piglet, and the total diarrhea score value was sorted from high to bottom, and the top 5 piglets with the highest total diarrhea score were selected as the total diarrhea score. Susceptible group (IS), the top 5 piglets with the lowest total diarrhea score served as the tolerant ...

Embodiment 2

[0064] Example 2. Overexpression of lncRNA ALDB-898 attenuates CPB2-induced inflammatory injury in IPEC-J2 cells

[0065] 1. Construction of LncRNA ALDB-898 overexpression vector

[0066] Step 1. Referring to the ALDB database of porcine lncRNA, IPEC-J2 cDNA was used as the template to amplify the full-length sequence of ALDB-898. PCR reaction system (20 μL): 2×Taq PCR Master Mix 10 μL, upstream primers (10 μmol / L) 1 μL, downstream primer (10 μmol / L) 1 μL, cDNA template 2 μL, RNA free water 6 μL. PCR reaction program: pre-denaturation at 95 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 60 °C for 30 s, extension at 72 °C for 60 s (31 cycles); extension at 72 °C for 10 min, and storage at 4 °C.

[0067] The upstream primer adds a Hind III (CAAGCTT) restriction site, and its nucleotide sequence is the sequence of SEQ ID NO.5.

[0068] The downstream primer added Xba I (CTCTAGA) restriction site, and its nucleotide sequence is the sequence of SEQ ID NO.6.

[0069]S...

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Abstract

The invention relates to the fields of genetic engineering and pig disease resistance genetics and breeding, and provides a lncRNA ALDB-898 sequence related to piglet bacterial diarrhea and an application thereof. The nucleotide sequence of the lncRNA ALDB-898 is such as SEQ ID NO. 1 shown. The present invention provides the fluorescent quantitative PCR detection primer pair of the lncRNA ALDB-898, as shown in SEQ ID NO. 2 and SEQ ID NO. 3), and confirms the objective existence of the lncRNA ALDB-898 by PCR, and it is in the C-type It was significantly downregulated in both C. perfringens-infected piglet ileum and in CPB2 toxin-treated intestinal epithelial cells (IPEC‑J2). The present invention constructs a lncRNA ALDB-898 over-expression vector, and finds that over-expression of lncRNA ALDB-898 can significantly inhibit the IPEC-J2 cytotoxicity and inflammation induced by CPB2 toxin, and provides a design target for the research and development of new prevention and control drugs for piglet bacterial diarrhea , and laid a technical foundation for the disease-resistant breeding of C. perfringens in piglets.

Description

technical field [0001] The invention relates to the fields of genetic engineering and pig disease resistance genetics and breeding, in particular to a lncRNA ALDB-898 related to piglet bacterial diarrhea and its application. Background technique [0002] Piglet bacterial diarrhea is a common intestinal disease in intensive pig production, which seriously affects the healthy development and economic benefits of the pig industry. Clostridium perfringens type C, as a common pathogen, is one of the main pathogenic bacteria causing diarrhea in piglets. Diarrhea caused by this bacteria is more likely to cause serious diseases due to its rapid onset and strong infectivity. CPB2 is one of the key pathogenic toxins produced by Clostridium perfringens type C, which can promote diarrhea and accelerate systemic infection through blood circulation. At present, the prevention and treatment of bacterial diarrhea in piglets only from the perspective of vaccines and antibiotics has limitati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/11C12Q1/6883C12Q1/6841C12N15/85A61K31/7105A61P1/12
CPCC12N15/113C12Q1/6883C12Q1/6841C12N15/85A61P1/12C12N2310/11C12Q2600/158C12Q2600/178C12N2800/107C12N2320/32C12Q2563/107C12Q2565/601
Inventor 高小莉滚双宝张生伟杨巧丽黄晓宇王鹏飞闫尊强
Owner GANSU AGRI UNIV
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