摘要
根据斑马鱼GDF9、BMP15基因的保守序列设计引物,采用RT-PCR方法扩增获得日本白鲫GDF9、BMP15基因片段。日本白鲫GDF9基因片段长778 bp,编码145个氨基酸;BMP15基因片段长811 bp,编码270个氨基酸。氨基酸序列分析表明日本白鲫GDF9与斑马鱼的同源性最高,为78%,与其他物种的同源性在58%～78%之间;日本白鲫BMP15与斑马鱼同源性最高,为80%,与其他物种的同源性在30%～80%之间。系统进化树分析显示,鱼类的GDF9基因独立聚成一支,其中日本白鲫的GDF9基因与斑马鱼的GDF9基因聚成一支;鱼类的BMP15基因独立聚成一支,其中日本白鲫的BMP15基因与斑马鱼的BMP15基因聚成一支。半定量PCR结果显示,BMP15 mRNA在脾脏、肝脏、鳃、脑、心脏、肌肉、肾脏、卵巢中均有表达,其中卵巢中表达量最高;GDF9 mRNA在肝脏、脑、心脏、肌肉、卵巢中均有表达,而在脾脏、鳃、肾脏中表达量极低或不表达,在卵巢中表达量最高,为进一步研究日本白鲫GDF9与BMP15基因的结构与功能奠定了基础。
According to zebrafish GDF9 and BMP15 gene conservative region of nucleotide sequence to design primers,the GDF9 and BMP15 gene fragment of Carassius curatus cuvieri was amplified by RT-PCR.The partial sequence of GDF9 is 778 bp and codes a protein of about 145 amino acids,the partial sequence of BMP15 is 811 bp and codes a protein of about 270 amino acids.The amino acid sequence comparison results showed that the GDF9 gene of Carassius curatus cuvieri shared 58%-78% identity with other species,and Carassius curatus cuvieri is most similar to zebrafish（78%）;BMP15 gene of Carassius curatus cuvieri shared 30%-80% identity with other species,and it is most similar to zebrafish（80%）.Phylogenetic tree of GDF9 and BMP15 gene generated by Neighbor Joining method suggested that all teleosts cluster together,and Carassius curatus cuvieri GDF9 is clustered closely with that of zebrafish,rainbow trout and European sea bass,and Carassius curatus cuvieri BMP15 is clustered closely with that of zebrafish and European sea bass.Then the expressions of different tissues including spleen,gill,liver,brain,heart,muscle,kidney and ovary were analyzed by semi-quantitative PCR.The results showed that the BMP15 gene of Carassius curatus cuvieri was expressed in all tissues and highest in ovary;the GDF9 gene of Carassius curatus cuvieri was expressed in liver,brain,heart,muscle and ovary,while highest in ovary and extremely low even no expression in spleen,gill and kidney.This study will be helpful for further understanding the structure and function of GDF9 and BMP15 gene.
出处
《上海海洋大学学报》
CAS
CSCD
北大核心
2012年第5期693-700,共8页
Journal of Shanghai Ocean University
基金
上海市重点学科水生生物学建设项目(S30701)
上海市教育委员会科研创新项目(11YZ151)
