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福建牡蛎17β-HSD基因的克隆及生殖周期表达
曾臻,倪健斌,谭强来,史博,余美舜,李伟杰,蔡广
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(1.厦门医学院海洋生物医药资源福建省高校工程研究中心,福建 厦门 361023;2. 厦门医学院天然化妆品福建省高校工程研究中心,福建 厦门 361023;3.厦门大学近海海洋环境科学国家重点实验室,福建 厦门 361102;4.集美大学水产学院,福建 厦门 361023)
摘要:
17β-羟基类固醇脱氢酶(17β-Hydroxysteroid dehydrogenase,17β-HSD)能够还原17-酮类固醇或者氧化17β-羟基类固醇,可以催化雄烯二醇和睾酮、雌酮和雌二醇、雄烯二酮和二氢睾酮之间的相互转化。本实验基于分子克隆的方法获得了福建牡蛎(Crassostrea angulata)类固醇激素合成相关酶基因17β-HSD的全长序列(全长1 149bp),可编码307个氨基酸。通过实时定量PCR(qRT-PCR)技术研究了该基因在福建牡蛎中的时空表达特征,该基因在牡蛎大多数组织中(性腺、闭壳肌、外套膜、鳃和内脏团)均有表达,其中在性腺中表达量最高(p<0.05),且在生殖周期的排放期出现较高的表达量。原位杂交结果显示该基因在卵巢滤泡细胞中表达。讨论了其在牡蛎性类固醇激素合成过程中的作用,为阐明牡蛎具有内源性类固醇激素的合成能力奠定基础,为进一步研究牡蛎生殖内分泌机理提供参考。
关键词:  海洋生物学  福建牡蛎  17β-HSD基因  生殖  性类固醇激素合成
DOI:
基金项目:国家自然科学基金资助项目(31702314);福建省卫生健康中青年骨干人才培养资助项目(2019-ZQNB-21);福建省中青年教师教育科研资助项目(JAT160585);海洋生物医药资源福建省高校工程研究中心(厦门医学院)开放课题资助项目(XMMC-MBS201901);天然化妆品福建省高校工程研究中心(厦门医学院)开放课题资助项目(XMMC-NC201901);厦门医学院自然科学研究资助项目(K2015-09)
Cloning of 17β-HSD gene and its characterization in the Fujian oyster, Crassostrea angulata, during gonad development
ZENG Zhen,NI Jian-bin,TAN Qiang-lai,SHI Bo,YU Mei-shun,LI Wei-jie,CAI Guang
(1. Engineering Research Center of Marine Biopharmaceutical Resource of Fujian Province,Xiamen Medical College, Xiamen 361023, China; 2. Engineering Research Center of Natural Cosmeceuticals College of Fujian Province, Xiamen Medical College,Xiamen Medical College, Xiamen 361023, China;3. State Key Laboratory of Marine Environmental Science, College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China; 4. Fisheries College of Jimei University, Xiamen 361023, China)
Abstract:
Crassostrea angulata is a major commercial species for the shellfish aquaculture in southern China. To investigate the reproductive endocrinology of mollusc, we studied the gene of enzymes involved in steroid metabolism in C. angulata with molecular biology technology. We obtained the fulllength cDNA 17β-HSD (1 149 bp) encoded peptides of 307 amino acids. 17β-HSD mRNA transcripts were expressed in most tissues of oyster with the highest level in gonad (p<0.05). The expression of 17β-HSD was lower during the gonad maturation of C. angulata, but higher in the initiation and partially spent stages (p<0.05). Furthermore, results obtained in situ hybridization indicated that 17β-HSD mRNA signals were detected in the follicle cell of ovary. Together with the in situ hybridization and qRT PCR results, it is suggested that 17β-HSD may play an important role in the regulation of gonad development based on steroidogenesis mechanism in C. angulata.
Key words:  marine biology  Crassostrea angulata  17β-HSD  reproduction  steroidogenesis

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