CAJ | 학술논문

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cDNA中克隆了fem-1c的cDNA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mRNA 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.
위진일보탐구어류성별결정적상관궤리, 증가대어류성공기인표체화공능적인식, 극륭반마어fem-1c기인병대기진행표체분석.연구채용RACE-PCR방법종반마어란소조직cDNA중극륭료fem-1c적cDNA전장서렬,기대소위2701 bp,편마618개안기산.생물신식학분석현시,반마어FEM-1C단백포함9개ANK결구역、2개TPR결구역화2개저복잡성구역, 여기타척추동물적FEM-1C단백서렬보수성교고.척추동물적fem-1c여tmed7、trim36등린근적4—5개기인구유보수적동선성관계.반정량RT-PCR실험결과현시반마어fem-1c재수정후17d개시표체, 병특이지표체우성체란소조직중.RNA원위잡교결과현시, fem-1c기인mRNA 정위우란소조직적Ⅰ기화Ⅱ기란모세포포질중.fem-1c 적시공표체특정암시기재반마어란소분화중구유중요작용.
Feminization-1 (fem-1) gene was first discovered in Caenorhabditis elegans, and itplays an important role in nematode sex determination. The mutation infem-1c could feminize the nematode sex differentiation. There are three conservative members offem-1 family in vertebrate, namely fem-1a,fem-1bandfem-1c. We cloned the entire cDNA sequence offem-1cfrom zebrafish ovary cDNA library using RACE-PCR. The full length of zebrafishfem-1c cDNA 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 betweenfem-1c and its neighborhood genes, especially the nearest 4—5 genes such as tmed7 andtrim36. We conducted the expression analysis in different tissues of adult zebrafish using RT-PCR and found thatfem-1c was exclusively expressed in the ovary. The expression analysis in the gonad of larvae at different development stages showed that the expression offem-1c began on the 17th day post-fertilization. Thein situ hybridization results showed thatfem-1c was expressed in cytoplasm at stage I andⅡ of oocyte. This specific expression pattern suggested thatfem-1c might play a role in zebrafish ovary development.