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Cell model having TLR4 immune response function and method for constructing same

A cell model, immune response technology, applied in the field of molecular biology, can solve problems such as the inability to study the function of TLR4

Active Publication Date: 2012-11-28
新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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Problems solved by technology

Therefore, HEK293 cannot be directly used for TLR4 function research

Method used

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  • Cell model having TLR4 immune response function and method for constructing same
  • Cell model having TLR4 immune response function and method for constructing same
  • Cell model having TLR4 immune response function and method for constructing same

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Embodiment Construction

[0019] The technology and features of the present invention will be described in detail below through specific examples, but these examples are only illustrative and not intended to limit the protection scope of the present invention.

[0020] (1) Primer design

[0021] According to the mRNA sequence (DQ319076) of bovine MD2 gene in GenBank, design primers for amplifying the CDS sequence of MD2 gene, the upstream primer is: ccg tga atc ​​atg ttt cca tttgtg ctt ttt tcc, the downstream primer is: gtt tgg gcc agg att ctc ctc gac gtc acc gca tgt tag cag act tcc tct gcc ctc tcc act gcc agc gta gtctgg gac gtc gta tgg gta gta att g. According to the mRNA sequence of the bovine CD14 gene in GenBank (AF141313.1), design primers for amplifying the CDS sequence of the CD14 gene. The upstream primer is: ccactc gag ctg act atg gtg tgc gtg ccc tac ctg ctg, downstream primers are: gtt acgcgt tgg gcc agg att ctc ctc gac gtc acc gca tgt tag cag act tcc tct gcc ctc tcc act gcc agc ...

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Abstract

The invention discloses a cell model having TLR4 immune response function and a method for constructing the cell model. The cell model and the method are characterized in that: three different bovine genes MD2, CD15 and TLR4 are connected and assembled to a vector by 2A sequence, the vector is delivered into a commercial cell line, and the constructed cell model has the function of LPS-mediated immune response. The cell model constructed in the invention and the construction method provide technical supports for the study of functions of the bovine TLR4 gene and the elucidation of the bovine TLR4 gene mechanism, therefore are valuable tool for studying bovine TLR4 gene functions.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular, the invention relates to a cell model with TLR4 immune response function and a construction method thereof. Background technique [0002] The natural immunity of eukaryotes is the first line of defense against the invasion of external pathogens. It mainly recognizes foreign pathogens through pattern recognition receptors, and then triggers a series of signal transduction, cytokine activation, and activation of the immune system. play an important role. Toll-like receptors (Toll Like Receptors, TLRs) are one of the model receptors that have been studied and discovered in recent years to recognize invading pathogens. TLRs can be the first to contact invading pathogens. By recognizing the pathogen-associated molecular pattern (PAMP) on the surface of pathogens, TLRs can activate and trigger the transcription and secretion of a series of chemokines and pro-inflammatory cytokines. For...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/85
Inventor 李文蓉白杰刘明军贺三刚张雪梅姚楠张宁玛依拉
Owner 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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