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Prokaryotic Expression, Antibody Preparation and Tissue Distribution of TLR10 Gene in Bactrian Camel (Camelus bactrianus) |
ZENG Wei-Wei, ZHANG Rui, XIE Dong-Xu, SUO Nan-Ji, WANG Ting-Wei, LIU Ke-Jiang, WANG Wen-Hui* |
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China |
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Abstract Toll-like receptors 10 (TLR10) plays an important role in the immune system by rapidly binding to ligands after pathogenic microorganisms enter the body and promoting the activation of various immune cells. However, the expression characteristics of TLR10 in different tissues and organs of the body need to be further explored. In this study, the TLR10 gene of Camelus bactrianus (GenBank No. XM_010962682.2) in NCBI was used to construct the prokaryotic expression plasmid pET-28a-TLR10. TLR10 recombinant protein was expressed in Escherichia coli, and purified protein was used to prepare TLR10 polyclonal antibody. ELISA and Western blot were used to detect the titer and specificity of the antibody. The results showed that the recombinant protein of bactrian camel was 50 kD in size and mainly expressed in the form of inclusion bodies. The antibody titer obtained after immunization with Oryctolagus cuniculus was 1∶32 000, and the expressed TLR10 protein could specifically bind to the antibody. TLR10 was mainly expressed in cardiomyocytes, hepatocytes and Kupffer cells, monocytes/macrophages in spleen, bronchial epithelial cells, typeⅡ alveolar cells and dust cells in lung and renal tubular epithelial cells in kidney. This study provides materials for further revealing the function of TLR10 protein and its biological role in the innate and adaptive immunity of bactrian camels.
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Received: 07 November 2022
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Corresponding Authors:
*wwh777@126.com
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