栉孔扇贝17β-HSD4基因的克隆和表达分析

Cloning and expression analysis of 17β-HSD4 gene in Chlamys farreri

  • 摘要: 性激素涉及动物的性别、性腺发育和生殖周期调控等功能,已知脊椎动物17β-羟基类固醇脱氢酶(17β-hydroxysteroid dehydrogenase,17β-HSD)在性激素合成和代谢中具有重要作用,而17β-HSD4主要参与雌二醇和雄烯二醇的灭活。本研究从栉孔扇贝(Chlamys farreri)转录组数据库中获得一个表达序列标签(EST),采用cDNA末端快速扩增(RACE)技术克隆得到1条全长为2417 bp cDNA序列,其开放阅读框为2223 bp,编码740个氨基酸。同源比对和系统进化分析结果显示其为17β-HSD4基因。半定量RT-PCR发现该基因在栉孔扇贝精巢、卵巢、肌肉、外套膜、鳃、肝胰腺和肾脏等各组织中均有表达,其中在肝胰腺和肾脏中表达量明显高于其他组织。qRT-PCR检测该基因在栉孔扇贝成熟期精巢的表达量显著高于增殖期和休止期的精巢和各时期卵巢的表达。由此推测栉孔扇贝多种组织中均可以合成17β-HSD4,并且可能参与精巢的成熟调控。

     

    Abstract: Sex hormones are involved in animal sex, gonad development and reproductive cycle regulation and other functions, it"s known that 17beta-hydroxysteroid dehydrogenase (17β-HSD4) had played an important role in synthesis and metabolism of the sex hormone. 17β-HSD4 mainly catalyzes to deactivate the biological activity of estradiol and androstenediol. In the present study, we got a partial expression sequence tag of 17β-HSD4 from the transcription group database of Chlamys farreri firstly, then a cDNA was cloned by using Rapid Amplification of cDNA End (RACE) methods, of which the full-length is 2217 bp, contains an open reading frame of 2223 bp and encodes a peptide of 740 amino acids. Homology analysis and phylogenetic tree revealed that it"s 17β-HSD4. The mRNA was detected widely in the testis, ovary, adductor muscle, mantle, gill, hepatopancreas and kidney, especially higher in hepatopancreas and kidney. The result of qRT-PCR showed the expression of 17β-HSD4 gene in testis at mature stage is obviously higher than the testis at proliferative stage and resting stage, and ovary of all stages. It could be concluded that 17β-HSD4 can be synthetized in many tissues of Chlamys farreri, also might regulate the maturity of testis.

     

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