稀有鮈鲫雄激素受体基因的克隆和内分泌干扰物对其表达的影响  

Full-length cDNA Cloning of Androgen Receptor Gene from Rare Minnow(Gobiocypris rarus) and Effect of Endocrine Disrupting Chemicals on Its mRNA Expression

作　　者：胡晓齐[1] 王晶晶[1] 王厚鹏[1] 吴婷婷[1] 秦芳[1] 任战军[1] 王在照[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《西北农业学报》2011年第2期8-14,共7页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：西北农林科技大学科研启动基金

摘　　要：采用RT-PCR和RACE法分离了稀有鮈鲫（Gobiocypris rarus）精巢雄激素受体基因（AR）的cDNA,其核苷酸序列3 130 bp,编码844个氨基酸。它的氨基酸序列与鲤科鱼类AR的同源性较高。AR基因在稀有鮈鲫的性腺、肝、脑、肠和肌肉等组织中均有表达,在雄性个体中精巢和肝脏的表达量最高,其他组织较低,而在雌性个体除肌肉中表达量较低外,其他组织均为中等水平的表达。0.01和0.1 nmol/L的乙炔基雌二醇暴露3 d后,能够分别非显著和显著地提高稀有鮈鲫幼鱼AR的mRNA表达,而1 nmol/L的乙炔基雌二醇则对其表达有下调的趋势,0.1~10 nmol/L的双酚A对其表达均有显著下调,0.01μmol/L壬基酚对其表达有显著下调,而0.1和1μmol/L壬基酚对其表达均有下调的趋势,因此不同种类内分泌干扰物及其不同暴露浓度对稀有鮈鲫AR的mRNA表达有不同影响。In the present study,RT-PCR and RACE（Rapid Amplification cDNA Ends） methods were used for the isolation of the cDNA of AR（androgen receptor） gene from testis of Gobiocypris rarus.Sequence analysis revealed a 3 130 bp cDNA encoding a protein of 844 amino acid residues.Phylogenetic analysis and multiple amino acids sequence alignment indicated the close relationship and high score similarity of rare minnow AR with ARs of other cyprinid species.AR mRNA expression was detected using semiquantitative RT-PCR in gonad,liver,brain,intestine,and muscle.AR mRNA was expressed at high levels in the testis and liver,low levels in the brain,intestine,muscle of male.However it was expressed at moderate levels in all the tissues except the muscle of female.Exposure to 0.01 and 0.1 nmol/L ethynylestradiol（EE2） for 3 days caused non-significant and significant increase of AR mRNA expression in rare minnow juveniles,respectively.On the contrary,1 nmol/L EE2 caused non-significant decrease in its expression.Bisphenol（BPA） from 0.1 to 10 nmol/L caused significant decrease of AR gene expression.0.01 μmol/L nonylphenol（NP） caused significant decrease of AR gene expression.While AR mRNA expression had down-regulation tendency in both 0.1 and 1 μmol/L NP exposed groups.Therefore AR gene expression was differentially modulated by various classes of EDCs and their different exposure concentrations.

关 键 词：稀有鮈鲫 雄激素受体 CDNA克隆 内分泌干扰物 实时定量PCR 

分 类 号：S917.4[农业科学—水产科学]