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Nucleotide sequence for detecting swine NLRP6 (nod-like receptor family pyrin domain-containing protein 6) and application of nucleotide sequence

A nucleotide sequence and sequence technology, which is applied in the detection of the nucleotide sequence and application field of porcine NLRP6, can solve the problems that have not been reported, and achieve the effect of high sensitivity and good specificity

Inactive Publication Date: 2016-08-24
TIANJIN INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] NLRP6 is a pathogen recognition receptor that has been studied more in the past two years, mainly focusing on the research on model animals and mice, and there are only a few reports on human research, but no research on pigs has been reported

Method used

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  • Nucleotide sequence for detecting swine NLRP6 (nod-like receptor family pyrin domain-containing protein 6) and application of nucleotide sequence
  • Nucleotide sequence for detecting swine NLRP6 (nod-like receptor family pyrin domain-containing protein 6) and application of nucleotide sequence
  • Nucleotide sequence for detecting swine NLRP6 (nod-like receptor family pyrin domain-containing protein 6) and application of nucleotide sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Relative Quantification of NLRP6 mRNA in Different Tissues of Pigs

[0050] (1) Porcine tissue sample processing: Use sterile scissors and tweezers to cut off about 1.0 g of the tissue sample to be tested, grind it thoroughly in a mortar, add 5 mL of PBS to mix, and then transfer the tissue suspension into a sterile Eppendorf tube , the numbers are reserved.

[0051] (2) RNA extraction: Use RNA extraction kits to extract RNA from different tissues, follow the kit’s operating instructions, and store the extracted RNA on ice for later use.

[0052] (3) RNA reverse transcription into cDNA: take out M-MLV (200 U / mL), 5´RT buffer, random primer (200 mM), RNase inhibitor (40 U / mL), dNTPs (2.5 mM each), MgCl 2 (25 mM), after thawing at room temperature, centrifuge at 2 000 rpm for 10 sec. Assuming that the number of PCR tubes required is n (n=number of samples + 1 tube of negative control), each test reaction system requires 10 mL of RT reaction solution, which contains 4 m...

Embodiment 2

[0060] Specificity test of detection method

[0061] Several common pathogens (comprising Escherichia coli, Salmonella, Staphylococcus and porcine reproductive and respiratory syndrome virus) in pig intestinal tissue and environment were detected with the method described in Example 1, and the results showed that the established method was consistent with pathogens. No cross-reactivity, good specificity ( figure 2 ).

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Abstract

The invention discloses a nucleotide sequence for detecting swine NLRP6 (nod-like receptor family pyrin domain-containing protein 6) and application of the nucleotide sequence. The nucleotide sequence for detecting the swine NLRP6 is shown as sequences SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3 in a sequence table. The sequences SEQ ID NO.1 and the SEQ ID NO.2 respectively represent sense primers and antisense primers for detecting the swine NLRP6, and the sequences SEQ ID NO.3 represent fluorescent probes for detecting the swine NLRP6. The invention further discloses a method for detecting the swine NLRP6 by the aid of the primers and the probes. The nucleotide sequence, the application and the method have the advantages that as shown by results, the method is good in specificity and high in sensitivity, expression of the swine NLRP6 in swine tissue and organ samples can be quickly and accurately detected, the swine NLRP6 can be monitored in real time when NLRP6 inflammasome is researched, inflammation occurrence procedures and development procedures can be timely understood and mastered, and the nucleotide sequence, the application and the method have important theoretical significance and practical significance.

Description

technical field [0001] The invention belongs to the field of animal medical technology research in animal husbandry, and relates to the nucleotide sequence and method for detecting porcine NLRP6 based on TaqMan hydrolysis probe fluorescent RT-PCR, which is suitable for porcine tissue samples, including stomach, small intestine, large intestine and other digestive tract tissues and liver Accurate detection of NLRP6 in tissues and organs such as spleen, lymph node, lung, etc., more specifically, it is used for the expression analysis of NLRP6 mRNA in pig tissues and organs, the research of inflammatory response mechanism and the research of intestinal barrier function. Background technique [0002] NOD-like receptors (NOD-like receptors, NLRs) are a large class of pathogenic pattern recognition receptors in innate immunity. The body relies on NLRs to recognize the signal of pathogenic infection, initiate an immune response, and play a role in resisting pathogenic microbial inf...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q2563/107C12Q2561/101C12Q2545/114
Inventor 李海花乔家运张全红朱琪赵向华王文杰
Owner TIANJIN INST OF ANIMAL HUSBANDRY & VETERINARY
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