| 摘要: |
| 利用简并引物扩增及RACE全长克隆技术, 克隆得到绿鳍马面鲀(Navodon septentrionalis)CYP19a基因cDNA全长序列。通过多序列比对, 发现具有芳香化酶特定保守序列, 包括一个I-螺旋区, 一个Ozol肽区, 一个亚铁血红素结合区域以及一个芳香化酶特异性结合区域。通过RT-PCR技术检测了其在绿鳍马面鲀成鱼各组织表达的情况, 发现其CYP19a基因只在卵巢中有表达; 同时也分析了其在不同卵巢发育期的表达情况, 发现CYP19a在卵黄发生后期表达量达到最高值, 卵巢退化吸收期表达量达到最低值。 |
| 关键词: 绿鳍马面鲀 P450芳香化酶 基因克隆 mRNA表达 |
| DOI：10.11693/hyhz201304037037 |
| 分类号: |
| 基金项目:山东省科技攻关计划课题资助, 2009GG10005017号。 |
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| THE GENE CLONING OF CYP19a AND ITS EXPRESSION CHARACTERIZATION ANALYSIS IN NAVODON SEPTENTRIONALIS |
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ZHANG Dong-Qian, WEN Hai-Shen, QIAN Kun, CHI Mei-Li, NI Meng, BU Yan, DING Yu-Xia
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College of Fisheries, Ocean University of China
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| Abstract: |
| Cytochrome P450s aromatizing enzyme is the key and rate-limiting enzyme to produce estradiol (E2). there are two different genes (CYP19a and CYP19b) encoding P450 aromatizing enzyme in teleost, and are both expressed in different tissues. The complete cDNA sequence of CYP19 in Navodon septentrionalis was obtained by the techniques of RT-PCR and rapid amplification of cDNA ends (RACE). The result of multiple amino acids alignment show that there was an I-coiled coil region, an Ozol’s peptide-binding region, a heme binding region, and a aromatizing enzyme specific binding region. The CYP19a mRNA expression in different tissues of the adult fish were analyzed by semi-quantitative PCR technique and the result suggest that it was expressed in ovary only. The result of CYP19a expression pattern in the ovary at different developmental stages show the highest expression before ovulation and lowest a after ovulation. |
| Key words: Navodon septentrionalis P450 aromatase gene cloning mRNA expression |
