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采用分子生物学方法成功克隆了虾夷马粪海胆Strongylocentrotus intermedius CYP17基因。该基因的cDNA全长为1 570 bp,其中开放阅读框1 482 bp,编码493个氨基酸;CYP17蛋白的相对分子质量约为55 540,等电点为8.068,信号肽断裂位点位于第18和第19个氨基酸残基之间;虾夷马粪海胆的CYP17氨基酸序列与紫球海胆S.purpuratus的同源性为97%,与光棘球海胆S.nudus的同源性为96%。用RT-PCR技术对CYP17基因在虾夷马粪海胆生殖腺不同发育时期的表达差异进行了分析。结果表明:在虾夷马粪海胆3个家系的雄性个体生殖腺中,CYP17基因在1-3家系的表达量最高,依次为10-3家系、6-2家系,而在雌性个体生殖腺中,CYP17基因在10-3家系的表达量最高,依次为1-3家系、6-2家系;在4-1家系海胆3个不同发育时期,CYP17基因在雄海胆性腺中的表达量随着日龄的增加而增加,而雌海胆性腺在发育第430天到第447天时表达量略有增加,发育到第512天时表达量明显增加。本研究表明,CYP17基因在海胆雌、雄个体生殖腺中转录水平上的表达差异明显,雄性的表达量明显高于雌性。
Strongylocentrotus intermedius CYP17 gene was successfully cloned by molecular biology method. The full-length cDNA of this gene was 1 570 bp in length, with an open reading frame of 1 482 bp encoding 493 amino acids. The relative molecular mass of CYP17 protein was about 55 540, the isoelectric point was 8.068, and the signal peptide cleavage site was at position 18 and The amino acid sequence of CYP17 shared 97% identity with S.purpuratus and 96% identity with S.nudus. The differences of CYP17 gene expression in gonad of Eriocheir japonica Semen from different developmental stages were analyzed by RT-PCR. The results showed that the CYP17 gene was most highly expressed in the 1-3 lines in the male individual gonads of 3 families of E. faecium, followed by 10-3 pedigrees and 6-2 pedigrees, while in female gonads, CYP17 The expression level of CYP17 gene in 10-3 pedigree was the highest, which was 1-3 pedigree and 6-2 pedigree. The expression level of CYP17 gene in male gonadal gonad at 3 different developmental stages of 4-1 family sea urchin increased with age While the expression level of gonads increased slightly from the 430th day to the 447th day after development, and the expression level increased significantly on the 512th day. This study showed that CYP17 gene expression in male and female gonads of the sea urchin significantly different levels of expression, male expression was significantly higher than the female.