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Specific primers, amplification procedures and quantitative kits for detecting the content of key molecules in the tlr2/4-mediated NF-κB signaling pathway

A signal pathway and specific technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., to reduce the use of funds, shorten the experimental time, and improve the accuracy

Active Publication Date: 2017-06-16
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on fluorescence quantification, the present invention uses a set of specific primers for fluorescence quantitative analysis to perform relative quantitative detection, and solves the current technical bottleneck of rapid detection of key molecules in the TLR2 / 4-mediated MyD88-dependent NF-κB signaling pathway

Method used

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  • Specific primers, amplification procedures and quantitative kits for detecting the content of key molecules in the tlr2/4-mediated NF-κB signaling pathway
  • Specific primers, amplification procedures and quantitative kits for detecting the content of key molecules in the tlr2/4-mediated NF-κB signaling pathway
  • Specific primers, amplification procedures and quantitative kits for detecting the content of key molecules in the tlr2/4-mediated NF-κB signaling pathway

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Experimental program
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Embodiment Construction

[0044] Using the primer sequence published by Genbank as a reference, download the known sequence from NCBI and use DNASTAR software for sequence comparison, select highly conserved and specific gene fragments for each target gene, and use Premier software to design relevant primers. '-3', the specific sequence is as follows:

[0045] Specific primers for quantitative PCR of TLR2, the sequences of which are shown in SEQ ID NO: 1 and SEQ ID NO: 2, and the sequences are complementary to a section of porcine TLR2 mRNA;

[0046] Specific primers for performing quantitative PCR on TLR4, the sequences of which are shown in SEQ ID NO: 3 and SEQ ID NO: 4, and the sequences are complementary to a section of porcine TLR4 mRNA;

[0047] Specific primers for performing quantitative PCR on TIRAP, the sequences of which are shown in SEQ ID NO: 5 and SEQ ID NO: 6, and the sequences are complementary to a section of porcine TIRAP mRNA;

[0048] Specific primers for quantitative PCR of IRAK1,...

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Abstract

The invention discloses a specific primer for detecting the content of key molecules in the NF-κB signaling pathway mediated by TLR2 / 4. The sequence is as follows: a specific primer for quantitative PCR of TLR2. The sequence is as follows: SEQ ID NO: 1, As shown in SEQ ID NO: 2, this sequence is complementary to a segment of porcine TLR2 mRNA. The beneficial effect of the present invention is that the TLR2 / 4-mediated differential expression of key molecules in the MyD88-dependent NF-κB signaling pathway plays a vital role in anti-virus research and is of great significance to the prevention and treatment of viral diseases. Through the design of the present invention, the reaction temperature of various primers is standardized, and the fragment length is also standardized, and all mRNA quantification can be completed under the same conditions, so that different experiments are comparable.

Description

technical field [0001] The invention relates to the field of biotechnology, and is a molecular biology method based on polymerase chain reaction (PCR), in particular to fluorescence quantitative polymerase chain reaction (Q-PCR). Background technique [0002] The NF-κB signaling pathway has a wide range of functions, especially in natural antiviral immune defense. According to the current research, the virus can induce the change of innate immune function by activating the NF-κB signaling pathway. At present, the study of virus pathogenicity or infection mechanism pays more attention to the change of host immune response during virus infection. After the virus infects the body, it is necessary to quickly and effectively detect the differential expression of key molecules in the NF-κB pathway for us to study the infection mechanism of the virus. Standardization, high cost and other reasons make experimenters face great difficulties. Contents of the invention [0003] The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 宋月魏战勇杨国宇徐端红索江华郑兰兰
Owner HENAN AGRICULTURAL UNIVERSITY