青虾，学名日本沼虾（Macrobrachium nipponense），是我国重要的淡水养殖经济虾类。随着生产规模的不断扩大，养殖青虾普遍出现性早熟，导致严重的“秋繁”现象和养殖过程中多代同堂、养殖密度增高、饲料用量增大而商品率下降，严重制约了青虾养殖业的健康发展。因此，开展青虾性腺发育相关基因的研究，不仅可为青虾遗传学研究积累背景资料，而且在控制性早熟甚至控制“秋繁”等方面具有广阔的应用前景。 雌性不育同源异型基因（female sterile homeotic, fsh）在果蝇中发现与雌性生殖发育相关，fsh基因的母源突变（maternal mutations）会引起体节缺失或是同源异生，而合子型的突变（zygotic mutations）将导致雌性不育或者是致死。该基因在甲壳动物中尚未有相关报道。从本实验室构建的青虾精巢文库中筛选到果蝇“雌性不育同源异型基因”的同源序列，采用快速扩增cDNA末端（rapid-amplification of cDNA ends, RACE）技术首次克隆了青虾“雌性不育同源异型基因”（Mnfsh）的全长cDNA序列。该cDNA序列全长2029 bp，包括1452 bp的开放阅读框（ORF）、361 bp的5'非编码区（UTR）和216 bp的3'UTR，其中3′末端发现含有2个典型的加尾信号AATAAA和10 bp的 Poly（A）尾巴。基因共编码483个氨基酸多肽前体，预测蛋白质分子量为52.641Da，理论等电点为8.78。氨基酸序列分析表明Mnfsh基因包含重要的Bromodomain结构域，没有信号肽及跨膜区域，为核内蛋白。Mnfsh基因与印度跳蚁（Harpegnathos saltator）、弓背蚁（Camponotus floridanus）和黑腹果蝇（Drosophila melanogaster）fsh基因比对分析显示，氨基酸序列相似性达到70%以上，一致性在50%-70%之间。在上述研究基础上，首次采用荧光定量PCR技术研究了Mnfsh基因的时空表达差异，结果表明，Mnfsh基因在成熟的未受精卵（O）中的表达量显著高于其他发育时期，随后在胚胎发育的早期迅速下降，在胚胎发育膜内蚤状幼体期（ZS期）、幼体发育期的第Ⅶ期幼体（LVII）和幼虾发育阶段的第一天（PL1）均有所恢复，达到表达小高峰。Mnfsh基因在6种成体组织中均有表达，表达水平依次为：卵巢>眼柄>肌肉>精巢>腹神经>心脏。 本研究首次克隆得到了青虾fsh基因的全长cDNA序列，对其结构特征进行了分析，并首次采用荧光定量PCR技术分析了该基因在青虾胚胎发育、幼体发育和幼虾发育的不同时期和成体组织的时空表达差异，这也是fsh基因在甲壳动物研究中的首次报道。结果预示Mnfsh基因可能为母本效应基因（maternal-effect gene），且与青虾发育过程中的组织分化和器官形成相关。本研究为深入开展青虾fsh基因功能等相关研究奠定了基础，并为青虾分子辅助育种工作积累背景资料，同时可为甲壳动物性腺发育研究提供借鉴。

Oriental river prawn, Macrobrachium nipponense, is an important shrimp species in freshwater aquaculture. However, sexual precocity is a common phenomenon with the extension of farming, leading to multigenerational together, high amount of feed and falling commodity rate, and heavily autumn propagation. These negative problems have seriously hampered the further development of oriental river prawn farming. Therefore, the researches to carry out the genes related with gonadal development in oriental river prawn will not only provide information for molecular assisted breeding, but also has important application value to control phenomena such as autumn propagation in practice. The maternal-effect gene female sterile homeotic (fsh) has been reported in Drosophila melanogaster, related to female reproduction and growth. According to the previous studies, zygotic mutations of fsh cause either lethality or female sterility, whereas maternal mutations cause segmental deletions and thoracic homeotic transformations. However, fsh was not studied and reported in crustaceans. In this study, a highly homologous fragment of fsh was identified from the testis of orinental river prawn Macrobrachium nipponense, named as Mnfsh and the full-length cDNA sequence was obtained for the first time by the combination of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The full-length cDNA sequence of fsh is 2029 bp, containing an open reading frame of 1452 bp, 361 bp of 5'-untranslated region (UTR), 462 bp of 3'-UTR with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail. The gene encodes a polypeptide of 483 amino acids with an estimated molecular mass of 52.641 kD and a theoretical pI of 8.78. The deduced amino acid sequence analysis showed that Mnfsh contains the Bromodomain region, whereas the gene does not have signal peptide and transmembrane region, implying it is the nuclear proteins. The amino acids of Mnfsh shares including 89%, 90% and 63% similarity to Harpegnathos saltator, Camponotus floridanus and Drosophila melanogaster, respectively, through the comparison with the amino acids of fsh in other insect. In addition, the spatial-temporal expression pattern of Mnfsh was detected by Quantitative real-time PCR (qRT-PCR) for the first time. According to the analysis results, the expression level reached the peak in matural ovum(O). The expression was slightly increased in zoea stage (ZS) during embryogenesis, the larvae VII (LVII) in the larvae and the 1st day of the post-larvae (PL1). Based on the expression pattern in adult tissues, the highest expression level was observed in ovary, followed by eyestalk, muscle, testis, abdominal ganglion and heart. It is the first time to clone the full length cDNA sequence of Mnfsh gene and analysed the structure of Mnfsh gene in this research. Besides, qRT-PCR technique was firstly used to detect the spatial-temporal expression pattern of Mnfsh gene during embryogenesis, larvae, post-larvae and expression pattern of adult tissues. The results strongly support that Mnfsh is the maternal-effect gene, revealing that Mnfsh is an arthropods fsh homologue and probably also played important roles in embryogenesis, organogenesis and morphological differentiation of oriental river prawn. This research could provide basic information for the function of Mnfsh gene and references for further studies in molecular assisted breeding, related to the gonadal development process of oriental river prawn, as well as in other crustacean species.