摘要
为从物种雌激素受体(ERα)敏感性差异的角度探索辽东湾野生梭鱼雌雄同体高发生率的一个可能原因,从梭鱼肝脏cDNA中扩增出长1113bp的梭鱼ERα(sERα)DEF区片段,并以此构建pGBT9-sERαDEF质粒,将其和pGAD424-TIF2质粒依次转入Y190酵母细胞中,建立了sERα物种特异性的酵母双杂交筛选系统.在此系统中测试了4种环境中主要的雌激素雌酮(E1)、17β-雌二醇(E2)、雌三醇(E3)及17α-炔雌醇(EE2)对梭鱼ERα的诱导活性,半数雌激素活性浓度(EC50)为3.7、2.4、1698.7、1.2nmol/L,分别是青鳉鱼ERα活性的4.6、2.3、3.4、3.3倍,说明梭鱼ERα对E1、E2、E3、EE2具有较高的敏感性,这为解释辽东湾梭鱼雌雄同体的高发生现象提供了重要线索.
To explore a possible reason why the wild So-iuy mullet of Liaodong Bay has high frequency of intersex from the point of view of specie-specific ERα binding sensitivity,a cDNA fragment of 1113 bp encoding So-iuy mullet （sERα） estrogen receptor DEF domain was amplified and used to construct plasmid pGBT9-sERα DEF.pGBT9-sERα DEF was transfected into Y190 yeast cells together with plasmid pGAD424-TIF2 for establishinig the sERα-specific in vitro system in yeast.Then we tested the sERα agonistic affinity of four major environmental estrogenic chemicals including estrone （E1）,17β-estrogen （E2）,estrodial （E3） and ethylestrodial （EE2）.Their half maximal concentrations （EC50）were calculated to be 3.7,2.4,1698.7,1.2 nmol/L,which are 4.6,2.3,3.4,3.3 times lower than those of Medaka ERα-binding activity,respectively.These results indicate that So-iuy mullet was more sensitive to E1,E2,E3,EE2 than medaka based on their ERα agonistic activity,providing an important clue to elucidate its remarkable occurence of intersex in Liaodong Bay.
出处
《中国环境科学》
EI
CAS
CSCD
北大核心
2010年第11期1490-1495,共6页
China Environmental Science
基金
国家自然科学基金资助项目(40632009)
国家"973"项目(2007CB407304)
关键词
ERΑ
梭鱼
酵母双杂交测试
物种敏感性
So-iuy mullet
estrogen receptor alpha
yeast two-hybrid
species-specific sensitivity
