虾夷马粪海胆CYP17基因的克隆及其表达差异  

Cloning and differential expression of CYP17 Gene in sea urchin Strongylocentrotus intermedius

作　　者：郭晓黎[1,2] 仇雪梅[1,2] 常亚青[1,2] 张伟杰[1,2] 崔军[2] 丛玉婷[2] 吴领知 刘洋 王秀利[1,2] 

机构地区：[1]大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连116023 [2]大连海洋大学水产与生命学院,辽宁大连116023

出　　处：《大连海洋大学学报》2013年第1期1-6,共6页Journal of Dalian Ocean University

基　　金：辽宁省自然科学基金资助项目(20102043);辽宁省"百千万人才工程"项目(2010921093);大连海洋大学科研项目(2012HYDX02)

摘　　要：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。In this study,CYP17 gene was cloned from sea urchin Strongylocentrotus intermedius by molecular biology methods.The results showed that the full length cDNA of CYP17 gene,derived relative molecular weight of about 55 540,and isoelectric point of 8.068,was found to be 1 570 bp including a full open reading-frame（ORF） of 1 482 bp encoding 493 amino acid residues.The predicted cleavage of the signal sequence to the putative mature peptide was located between the 18th and 19th amino acid residues.The amino sequence analysis indicated that the CYP17 of S.intermedius has high homology with S.purpuratus（97%）,and S.nudus（96%）.The CYP17 gene expression at transcription level was analyzed in different developmental stages of the gonad of S.intermedius by real-time RT-PCR.The results showed that the maximal expression level of CYP17 gene was in family 1-3,higher in family 10-3 and the minimal in family 6-2 in the testis among the three S.intermedius families.The CYP17 gene expression level was increased with the testis development in family 4-1.However,the expression level of CYP17 gene in the ovary was increased slightly from 430 d to 447 d,and increased obviously when the gonads develop to 512 d.The findings indicate that the expression of CYP17 gene in the sea urchin gonad showed significant difference between female and male,the significantly higher expression level in male than in female.

关 键 词：虾夷马粪海胆 CYP17基因 克隆 基因表达 

分 类 号：S917[农业科学—水产科学]