鲈鱼（欧洲鲈）肾脏中一种邻苯二甲酸酯诱导型CYP4基因（CYP4T2）的分子克隆
Molecular cloning of a phthalate-inducible CYP4 gene (CYP4T2) in kidney from the sea bass, Dicentrarchus labrax.
作者信息
Sabourault C, Bergé J, Lafaurie M, Girard J P, Amichot M
机构信息
INRA-IFR 38 "Réponse des Organismes aux Stress Physico-Chimiques et Biologiques,", 123 Bd Francis Meilland, Antibes Cedex, 06606, France.
出版信息
Biochem Biophys Res Commun. 1998 Oct 9;251(1):213-9. doi: 10.1006/bbrc.1998.9429.
A full-length cDNA sequence was isolated from kidney total RNA of di(2-ethylhexyl) phthalate-treated sea bass by reverse-transcriptase polymerase chain reaction and then rapid amplification of cDNA ends. The deduced amino acid sequence, which has been named CYP4T2, shared 69 and 54.4% amino acid identity with rainbow trout CYP4T1 and rat CYP4B1, respectively. RNA blot analysis using the CYP4T2 cDNA as a probe indicated that the mRNA was rather abundant in kidney, and less so in liver, small intestine, and brain. Treatment of sea bass with peroxisome proliferators showed marked tissue-specific induction. CYP4 inducers clofibrate, di(2-ethylhexyl) phthalate (DEHP), and 2,4-dichlorophenoxy acetic acid (2,4-D) were administered by intraperitoneal injection. The strongest induction was found in kidney after a DEHP treatment (6.5-fold) or a 2,4-D treatment (9-fold), while no induction was observed in liver.
通过逆转录聚合酶链反应从邻苯二甲酸二（2-乙基己基）酯处理的海鲈鱼肾脏总RNA中分离出全长cDNA序列，然后进行cDNA末端快速扩增。推导的氨基酸序列被命名为CYP4T2，与虹鳟鱼CYP4T1和大鼠CYP4B1的氨基酸同一性分别为69%和54.4%。以CYP4T2 cDNA为探针的RNA印迹分析表明，该mRNA在肾脏中相当丰富，而在肝脏、小肠和大脑中较少。用过氧化物酶体增殖剂处理海鲈鱼显示出明显的组织特异性诱导。通过腹腔注射给予CYP诱导剂氯贝丁酯、邻苯二甲酸二（2-乙基己基）酯（DEHP）和2,4-二氯苯氧基乙酸（2,4-D）。在DEHP处理（6.5倍）或2,4-D处理（9倍）后，在肾脏中发现最强的诱导，而在肝脏中未观察到诱导。
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