Duck TLR2 extracellular region polyclonal antibody as well as preparation method and application thereof

A polyclonal antibody, extracellular domain technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve problems such as unreported, and achieve the effect of good specificity and high purity

Inactive Publication Date: 2018-11-23
JIANGSU INST OF POULTRY SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] In other species, especially model animals, there have been antibody preparation and functional studies of various TLRs family members, some of which have formed commercial products, but this research has not been reported in poultry TLRs research, especially waterfowl research

Method used

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  • Duck TLR2 extracellular region polyclonal antibody as well as preparation method and application thereof
  • Duck TLR2 extracellular region polyclonal antibody as well as preparation method and application thereof
  • Duck TLR2 extracellular region polyclonal antibody as well as preparation method and application thereof

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Embodiment Construction

[0025] The present invention will be explained in detail below in conjunction with the accompanying drawings and embodiments.

[0026] The preparation method of duck TLR2 extracellular region polyclonal antibody specifically comprises the following steps:

[0027] (1) Fragment amplification of the extracellular region of duck TLR2

[0028] Using Predict Protein software to analyze the structure of amino acids encoded by duck HQ166194.1 (16S ribosomal RNA coding gene sequence), infer the range of active extracellular region, and screen the region to be expressed in recombinant protein.

[0029] The present invention is based on HQ166194.1, and after comparison and analysis with the duck TLR2 coding sequence published by NCBI, the predict protein software is used for analysis. The gene encodes 693 amino acids, 1-25aa is the signal peptide sequence, and the amino acid display is 2 times Transmembrane, 558-577, 598-617 is the transmembrane region; 648-693aa is the TIR region, and...

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Abstract

The invention discloses a duck TLR2 extracellular region polyclonal antibody as well as a preparation method and application thereof. The method comprises the following steps: (1) artificially synthesizing a TLR2 extracellular region sequence; (2) constructing the TLR2 extracellular region sequence to an expression carrier pET28a and screening to obtain recombinant plasmid pET28a-TLR2; (3) converting recombinant plasmid pET28a-TLR2 to Escherichia coli BL21 and expressing TLR2-His recombinant protein in the Escherichia coli BL21; (4) utilizing the TLR2-His recombinant protein to immunize domestic rabbits and separating immunized domestic rabbit serum to obtain the duck TLR2 extracellular region polyclonal antibody. The serum titer of the prepared polyclonal antibody disclosed by the invention exceeds 1 to 50000, the polyclonal antibody has the characteristics of good specificity and high purity, recombinant protein and tissue endogenous TLR2 can be specifically detected, and the prepared polyclonal antibody provides advantageous conditions for further researching biological functions of TLR2.

Description

technical field [0001] The invention relates to a preparation method of an antibody, in particular to a duck TLR2 extracellular region polyclonal antibody and its preparation method and application. Background technique [0002] TLR2 is an important member of the TLRs family. It is mainly expressed on the surface of mononuclear macrophages. Therefore, TLR2 is not only a key molecule connecting innate immunity and acquired immunity, but also plays an important role in the process of non-infectious tissue damage and tissue repair. [0003] At present, the structure and function of TLR2 in mammals and non-mammalian vertebrates such as humans, mice, cattle, pigs and chickens have been extensively studied. It has been proven that the full-length CDS of human, mouse and bovine TLR2 genes is 2355bp, encoding 784aa, including 54aa of C-terminal leucine-rich repeats (LRRs) and 145aa of TIR homology region; the full-length CDS of pig TLR2 gene is 2358bp, It encodes 785aa, of which t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06C07K14/47C12N15/12C12N15/70A61K39/395A61K47/68A61P37/00
CPCA61K2039/505A61K47/6849A61P37/00C07K14/47C07K16/065C07K16/18C07K2317/33C12N15/70
Inventor 朱春红李慧芳陶志云徐文娟宋卫涛刘宏祥章双杰
Owner JIANGSU INST OF POULTRY SCI
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