Peptide characterization and immunolocalization of Echinococcus granulosus aquaporin 9
TAN Qing-qing1, LI Kai1, ZHANG Jing1, 2, GAO Jian3, WU Hong-ye1, FAN Jun-jie1, LU He1, 2, YE Bin1, 2
1.Department of Pathogenic Biology, Chongqing Medical University, Chongqing 400016, China;
2.Research Center for Molecular Medicine and Tumor, Chongqing Medical University, Chongqing 400016, China;
3.Department of Parasitology, Xinjiang Medical University, Urumqi 830011, China
Abstract：A specific polypeptide antibody of Echinococcus granulosus aquaporin 9 (EgAQP9) was developed and used to detect the tissue distribution of this protein in the germinal cells, protoscoleces and hydatid cyst walls of Echinococcus granulosus. The B cell antigen polypeptide was designed on the basis of the amino acid sequence of EgAQP9, and the polyclonal antibodies was prepared by using the synthesized polypeptide coupled with KLH as an immunogen to immunize New Zealand rabbits. The multi-antibody titer was determined with ELISA, the immunoactivity was analyzed with western blotting, the subcellular localization of EgAQP9 was determined with immunofluorescence assays, and the tissue localization of EgAQP9 was determined with immunohistochemistry assays. The polyclonal antibody was successfully prepared, with a titer as high as 256 000, according to ELISA. Western blotting revealed that the prepared antibody allowed for specific identification of bands at 36 kDa in natural E. granulosus; this mass was similar to the predicted relative molecular weight of EgAQP9. Furthermore, immunofluorescence assays indicated that EgAQP9 was distributed in the cytoplasm and membranes of the germinal cells of hydatids, and immunohistochemistry assays showed that the protein was mainly distributed at the surfaces of the protoscoleces and in the germinal layers of the hydatid cysts. The prepared EgAQP9 rabbit-derived polyclonal antibody was used to determine the distribution of EgAQP9 protein in the germinal cells, protoscoleces and cyst walls of E. granulosus hydatids.
谭青青, 李锴, 张静, 高剑, 吴宏烨, 范俊杰, 陆合, 叶彬. 细粒棘球绦虫水通道蛋白9的多肽特征及免疫定位研究[J]. 中国人兽共患病学报, 2021, 37(1): 45-52.
TAN Qing-qing, LI Kai, ZHANG Jing, GAO Jian, WU Hong-ye, FAN Jun-jie, LU He, YE Bin. Peptide characterization and immunolocalization of Echinococcus granulosus aquaporin 9. Chinese Journal of Zoonoses, 2021, 37(1): 45-52.
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