In this study, we used bioinformatics software to analyze the TLR1 (Toll-like receptors 1) gene and used a prokaryotic expression system to inducibly express TLR1 recombinant plasmids into TLR1 protein. Rabbit polyclonal antibactrian camel TLR1 antibody was generated using the expressed recombinant TLR1 protein, and the expression of TLR1 in organs including bactrian camel heart, liver, spleen, lung, and kidney was detected by HE staining coupled with immunohistochemical staining of SABC using the prepared TLR1 polyclonal antibody. The size of the protein of interest obtained in this study was 62 kDa, which was the same as predicted by bioinformatics software. TLR1 recombinant protein was mainly expressed in the form of inclusion bodies. In this study, the titer of the TLR1 antibody was determined by indirect ELISA, which showed that the antibody titer result was 1∶ 64 000, and the specificity of the TLR1 antibody was confirmed by Western blot analysis. The results of the present trial of HE staining with immunohistochemical staining of SABC showed that TLR1 was upregulated in cardiomyocytes of bactrian camel hearts, Kupffer cells of the liver, monocytes/macrophages of the spleen, the respiratory bronchiolar epithelium, and type Ⅰ and type Ⅱ alveolar cells of the lung. A positive reaction appeared on the epithelial cells of the distal convoluted tubule of the kidney. In conclusion, the rabbit anti-bactrian camel TLR1 polyclonal antibody has good specificity and can be used for immunohistochemical detection of TLR1 and provides a basis for further investigation of the function of TLR1 in different tissues and organs of bactrian camels.
XIE Dongxu
,
ZHANG Rui
,
Suonanji
,
LIU Kejiang
,
WANG Tingwei
,
WANG Wenhui
. Prokaryotic expression system construction, antibody preparation and tissues distribution of bactrian camel TLR1[J]. ACTA THERIOLOGICA SINICA, 2023
, 43(6)
: 745
-752
.
DOI: 10.16829/j.slxb.150780
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