Pig circRNA sequence, application thereof and cyclization identification method

A sequence and looping technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of less research on porcine circRNA

Inactive Publication Date: 2021-12-28
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the research on the function of circRNA mainly focuses on the interaction with miRNA, protein regulation and cancer and other diseases, but there are relatively few studies on pig circRNA

Method used

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  • Pig circRNA sequence, application thereof and cyclization identification method
  • Pig circRNA sequence, application thereof and cyclization identification method
  • Pig circRNA sequence, application thereof and cyclization identification method

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Experimental program
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Effect test

Embodiment Construction

[0037] 1. Materials and methods

[0038] 1.1 RNA and genomic DNA (gDNA) extraction

[0039] In this study, the Trizol extraction kit (Invitrogen, USA) was used to extract total RNA from porcine small intestinal epithelial cells (IPEC-J2). The cells were lysed by adding the lysate according to the instructions, and then the binding solution was added to mix and pass through the column. The column was washed twice, and the RNA was finally eluted and tested for RNA purity, concentration, and integrity. Genomic DNA (gDNA) extraction was performed according to the instructions of the kit (Sangon Biotech, China).

[0040] 1.2 RNase R treatment, cDNA synthesis and PCR amplification

[0041] The same aliquot of RNA (2 μg) samples was incubated with 3 U / μg RNase R (Epicenter Technologies, USA) for 30 minutes at 37 °C, and then processed by RNeasy MinElute purification kit (Qiagen, Germany). Operate according to the instructions of the reverse transcription kit, and use RNA as a templa...

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Abstract

The invention is based on the most common intestinal inflammatory disease, namely piglet bacterial diarrhea (PDW), wherein the F18 Escherichia coli (E. coli F18) is a main pathogen causing weaned piglet bacterial diarrhea, and regulation mechanisms of local variety pigs to the resistance of the E. coli F18 are to be systematically clarified. In the early stage, a key candidate RNA-circRNA-27388 for escherichia coli F18 infection regulation is screened out through omics sequencing, as a circRNA-27388 body gene is FUT8, the circRNA-27388 body gene is named as circ-FUT8, on the basis, the circ-FUT8 sequence is subjected to cyclization identification, and a foundation is laid for follow-up verification of circRNA functions and the regulation mechanisms.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the verification of the circle formation of circ_FUT8 in piglet bacterial diarrhea (PDW) circRNA and its application. Background technique [0002] Escherichia coli F18 is one of the important pathogenic bacteria that cause bacterial diarrhea in weaned piglets in today's pig industry. Symptoms such as edema and diarrhea occur. Therefore, whether weaned piglets can resist the infection of E. coli F18 mainly depends on whether the E. coli F18 receptor of the small intestinal mucosal epithelial cells expresses. Foreign studies have found that type 1 H antigen, which participates in the composition of ABH blood group antigen, is the smallest epitope of the adhesion receptor of F18 Escherichia coli, mainly catalyzed by α-(1, 2) fucosyltransferase (FUT2 enzyme), and FUT2 enzyme The functional activity of FUT1 is jointly regulated by the FUT1 and FUT2 genes; foreign scholars have found th...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6888C12Q1/6851
CPCC12N15/113C12Q1/6888C12Q1/6851C12N2310/532C12Q2600/124C12Q2600/158C12Q2600/178C12Q2531/113C12Q2521/107C12Q2521/327C12Q2527/125C12Q2525/307
Inventor 吴正常许写周荣任战士包文斌吴圣龙
Owner YANGZHOU UNIV
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