摘要
为进一步探究鱼类性别决定的相关机理,增加对鱼类性控基因表达和功能的认识,克隆斑马鱼fem-1c基因并对其进行表达分析。研究采用RACE-PCR方法从斑马鱼卵巢组织c DNA中克隆了fem-1c的c DNA全长序列,其大小为2701 bp,编码618个氨基酸。生物信息学分析显示,斑马鱼FEM-1C蛋白包含9个ANK结构域、2个TPR结构域和2个低复杂性区域,与其他脊椎动物的FEM-1C蛋白序列保守性较高。脊椎动物的fem-1c与tmed7、trim36等邻近的4—5个基因具有保守的同线性关系。半定量RT-PCR实验结果显示斑马鱼fem-1c在受精后17d开始表达,并特异地表达于成体卵巢组织中。RNA原位杂交结果显示,fem-1c基因m RNA定位于卵巢组织的Ⅰ期和Ⅱ期卵母细胞胞质中。fem-1c的时空表达特征暗示其在斑马鱼卵巢分化中具有重要作用。
Feminization-1（fem-1） gene was first discovered in Caenorhabditis elegans, and it plays an important role in nematode sex determination. The mutation in fem-1c could feminize the nematode sex differentiation. There are three conservative members of fem-1 family in vertebrate, namely fem-1a, fem-1b and fem-1c. We cloned the entire c DNA sequence of fem-1c from zebrafish ovary c DNA library using RACE-PCR. The full length of zebrafish fem-1c c DNA was 2701 bp, encoding a 618-amino acid protein. We used online bioinformatic software and predicted that the FEM-1C protein contained 9 ANK domains, 2 TPR domains and 2 low complexity regions. Zebrafish FEM-1C shared a highly conservative protein sequence with its homologues in other vertebrates. The sequence analysis also revealed that in vertebrates there was a conservative synteny between fem-1c and its neighborhood genes, especially the nearest 4—5 genes such as tmed7 and trim36. We conducted the expression analysis in different tissues of adult zebrafish using RT-PCR and found that fem-1c was exclusively expressed in the ovary. The expression analysis in the gonad of larvae at different development stages showed that the expression of fem-1c began on the 17 th day post-fertilization. The in situ hybridization results showed that fem-1c was expressed in cytoplasm at stage I and Ⅱ of oocyte. This specific expression pattern suggested that fem-1c might play a role in zebrafish ovary development.
出处
《水生生物学报》
CAS
CSCD
北大核心
2015年第3期459-467,共9页
Acta Hydrobiologica Sinica
基金
国家973计划(2010CB126301)
现代农业产业技术体系建设专项资金(NYCYTX-49)资助
关键词
fem-1c
卵巢分化
原位杂交
斑马鱼
fem-lc
Ovary differentiation
In situ hybridization
Zebrafish
