【摘要】 目的:研究黑脊倒刺鲃scnanos的核苷酸序列及其在配子发生中的空间表达特征,以期为鱼类生殖发育研究提供一个有效的分子标记。方法:RT-PCR法首次克隆出scnanos基因,并检测其在不同组织中的表达水平,进而原位杂交法检测其在性腺中的表达水平。结果:scnanos开放阅读框长456bp,编码152个氨基酸。其具有较高的保守性,与鲤鱼、斑马鱼、家蚕、大熊猫、紫色球海胆的相似性分别为81%、63%、52%、50%、44%;scnanos仅在性腺表达,精子发生中,scnanos仅在精原细胞和精母细胞中表达。卵子发生的各时期都发现有scnanos的表达,且杂交信号的强度都随着配子发生的进行逐渐减弱。结论:该研究得到的scnanos是斑马鱼nanos的同源基因,而且此基因适合作为黑脊倒刺鲃及其它鱼类生殖发育研究的有效分子标记基因。更多还原

【Abstract】 Objective:The nucleotide sequence of nanos homolog from Spinibarbus caldwelli and its expression pattern in gametogenesis were studied in order to search for a useful molecular marker for fish developmental research.Method:Nanos homolog from Spinibarbus caldwelli was successfully cloned for the first time and named as scnanos(Spinibarbus caldwelli nanos).Its expression levels in different tissues or gonads were assayed by the method of RT-PCR and in situ RNA hybridization.Result:Scnanos contains an open reading frame(456 bp),which encodes 152 amino acids.Based on the aligning result,Spinibarbus caldwelli nanos was highly conserved and showed high similarity to that of Cyprinus carpio(81%),Danio rerio(63%),Bombyx mori(52%),Ailuropoda melanoleuca(50%),Strongylocentrotus purpuratus(44%).RT-PCR showed that scnanos was expressed specifically in Spinibarbus caldwelli gonad tissue instead of other tissues.In situ RNA hybridization analysis result was as follows:During spermatogenesis,scnanos was transcripted only in spermatogonia and spermatocytes.During oogenesis,scnanos was transcripted in every phase from oogonia to oocytes.The earlier phase of oogenesis,the more scnanos mRNA molecules were transcripted.Conclusion:Scnanos is the Spinibarbus caldwelli homolog of the zebrafish nanos.Also,it could be used as a useful marker for development research of fish germ cell.更多还原