Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii
Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii
复制标题
罗氏沼虾卵巢特异性基因 Mrfem-1 的分子特征以及 siRNA 介导的针对 Mrfem-1 的调控
DOI：
10.1016/j.gene.2020.144891
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发表时间：
2020
期刊：
影响因子：
3.5
通讯作者：
Qiu Gaofeng
中科院分区：
文献类型：
--
作者：
Zhou Lingxia;Liu Xue;Ye Baoqing;Liu Yun;Tan Shuangpei;Ma Keyi;Qiu Gaofeng
fem-1基因在秀丽隐杆线虫性别决定/分化过程中起重要作用。然而，在甲壳类动物中fem-1的研究报道很少。本研究首次从罗氏沼虾（Macrobrachium rosenbertus）中分离到afem-1基因（Mrfem-1）。Mrfem-1的cDNA全长为2607 bp，包含一个开放阅读框，编码615个氨基酸，有8个锚蛋白重复序列。该基因已在GenBank中登录，登录号为MT 160093。RT-PCR结果表明，Mrfem-1在卵巢中特异表达。随着卵巢成熟，Mrfem-1的表达量逐渐增加，在卵黄发生期达到最高峰，原位杂交结果显示，卵黄发生前期阳性信号集中在细胞质中，卵黄发生期阳性信号散在分布于细胞质和卵泡细胞中，提示Mrfem-1可能与卵巢成熟有关。此外，还发现了两个靶向Mrfem-1的siRNA，并在体外验证了它们的有效性。Mrfem-1的研究结果将有助于我们更好地了解fem家族，并为后续研究siRNA介导的卵巢发育调控提供新的资源。罗森伯托。
Characterized by ankyrin repeat motifs, thefeminization-1(fem-1) gene plays an essential role in sex determination/differentiation inCaenorhabditis elegans. However, there are only a few reports onfem-1in crustaceans. In this study, afem-1gene (Mrfem-1) was first isolated from the giant freshwater prawnMacrobrachium rosenbergii. The full-length cDNA ofMrfem-1was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results,Mrfem-1was exclusively expressed in the ovary. The expression level ofMrfem-1had increased with ovarian maturation and reached the highest peak at vitellogenic stage.In situhybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting thatMrfem-1might be associated with ovarian maturation. Moreover, two effective siRNAs targetingMrfem-1were found and their effectiveness verifiedin vitro. These results onMrfem-1will help us to better understand thefemfamily and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development inM. rosenbergii.