水产学报2018，Vol.42Issue(1)：39-47,9.DOI:10.11964/jfc.20161110605
三疣梭子蟹B型清道夫受体蛋白的原核表达及细胞、组织分布
Prokaryotic expression, cell and tissue distribution of the class B scavenger receptor in Portunus trituberculatus
摘要
Abstract
Class B scavenger receptors are a subclass of scavenger receptors family that can recognize a large rep-ertoire of ligands, and they have been shown to have a wide range of functions in lipids metabolism and immune defence. To better understand the functions of class B scavenger receptors in Portunus trituberculatus (Pt-SRB), the extracellular loop of Pt-SRB was sequenced and subcloned into the expression vector, and the expressed re-combinant rPt-SRB protein was obtained in present study. Then the rPt-SRB was purified by nickel-nitrilotriacetic acid chromatography, and the polyclonal antibody against rPt-SRB was obtained from an immunized rat using a conventional method. Results of SDS-PAGE showed that the recombinant protein had a molecular mass of 49.18 ku and was predominantly expressed in the insoluble fraction. Moreover, Western-blot analysis revealed that anti-serum could specifically react with the rPt-SRB. Furthermore, immunofluorescent staining technique was used to determine the tissue distribution and the cellular localization of Pt-SRB in this study. The results showed that the Pt-SRB were expressed in the tissues including intestine, hepatopancreas, gills, heart and muscle, but the Pt-SRB were located in different tissue structures. The results displayed that the green fluorescent signals were mainly loc-ated in the connective tissue of digestive tract and gland, epithelial cells of the gill and hepatopancreatic tubules. In the crab hemocytes, the green fluorescent signals were located in the membrane and cytoplasm, but not in the nuc-leus, which suggested that Pt-SRB was localized both in membrane and cytoplasm in the crab hemocytes. Our findings will help us to better understand potential functions of Pt-SRB.关键词
三疣梭子蟹/B型清道夫受体/原核表达/组织分布Key words
Portunus trituberculatus/Class B scavenger receptor/prokaryotic expression/tissue distribution分类
生物科学引用本文复制引用
杨宁,周素明,王国良,刘顺,李猛..三疣梭子蟹B型清道夫受体蛋白的原核表达及细胞、组织分布[J].水产学报,2018,42(1):39-47,9.基金项目
宁波市海洋蟹类产业科技创新团队项目(2011B81003) （2011B81003）
宁波大学"水产"浙江省重中之重开放基金(xkzsc1404) （xkzsc1404）
Ningbo Municipal Innovative Research Team (2011B81003) （2011B81003）
Open Fund of Zhejiang Provin-cial Top Key Discipline of Aquaculture of Ningbo University (xkzsc1404) （xkzsc1404）
