食蚊鱼P-糖蛋白基因克隆及三氯生对其mRNA表达的影响Cloning of P-Glycoprotein Gene and Effects of Triclosan on Its mRNA Expression in Mosquitofish(Gambusia affinis)
宋晓红;陆爱金;杜士林;陈双凤;陈旺光;梁延鹏;黄亮亮;曾鸿鹄;
摘要(Abstract):
三氯生(TCS)是一种广谱高效抗菌剂,在水环境和生物体内均不同程度检出,对水生生物具有潜在风险。P-糖蛋白(Pgp)是生物体多型异源物质抗性防御系统中重要的"膜解毒蛋白",对水生生物体内的有毒物质和代谢产物具有重要的外排和转运作用。为探究P-gp在鱼类免疫中的作用,克隆了食蚊鱼(Gambusia affinis) P-gp基因的c DNA,检测了不同浓度(50、100和150μg·L~(-1)) TCS暴露12 h、1 d、3 d、5 d、7 d和9 d后,P-gpmRNA相对表达量的变化。实验获得的食蚊鱼P-gpcDNA共5 452bp,编码1 294个氨基酸,具有ABC转运蛋白家族典型的跨膜结构、功能区域和作用位点,与其他鳉形目的鱼类P-gp氨基酸序列同源性较高。TCS对P-gpmRNA表达的影响呈现倒"U"型的时间-剂量效应,50μg·L~(-1)和100μg·L~(-1)TCS胁迫后P-gp表达量先升高后下降,100μg·L~(-1)暴露组表达高峰在暴露1 d时,50μg·L~(-1)TCS暴露组表达高峰延迟至3 d,而150μg·L~(-1)暴露组Pgp表达无显著变化。结果表明,P-gp基因参与了TCS胁迫的解毒过程,有助于食蚊鱼抵抗外源污染物的毒性作用。
关键词(KeyWords): 三氯生;食蚊鱼;P-糖蛋白基因;cDNA克隆;mRNA表达
基金项目(Foundation): 国家自然科学基金资助项目(51578171);; 广西科技计划项目(桂科AD18126018);; 广西自然科学基金资助项目(2018GXNSFAA281022)
作者(Author): 宋晓红;陆爱金;杜士林;陈双凤;陈旺光;梁延鹏;黄亮亮;曾鸿鹄;
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