半滑舌鳎含sox9基因BAC克隆的筛选及BAC-FISH定位

Screening and BAC-FISH Locating of the sox9 Gene BAC Clone of Half-Smooth Tongue Sole Cynoglossus semilaevis

中文关键词: 半滑舌鳎  sox9基因  BAC文库  荧光原位杂交  染色体

英文关键词: Cynoglossus semilaevis  sox9 gene  BAC library  fluorescence in situ hybridization (FISH)  chromosome

中文摘要:

      以本实验室构建的半滑舌鳎BAC文库为基础，通过对其进行有序混合，构建了两步PCR筛选体系；对性别决定相关的sox9基因的序列设计特异性引物，筛选获得阳性单一BAC克隆。利用BAC-FISH技术将此包含sox9基因的BAC克隆定位在半滑舌鳎染色体上。结果显示，sox9基因在雌、雄半滑舌鳎的一对常染色体的长臂上分别存在2个杂交信号位点，信号稳定且特异。研究证实了该文库筛选体系的有效性；首次实现了对性别相关的sox9基因在半滑舌鳎染色体上的定位，其结果为sox9基因深入研究，以揭示其参与鱼类性别控制的机制以及半滑舌鳎染色体鉴别等工作的进行提供了重要的基础。

英文摘要:

      A two-step-PCR screening system of the half-smooth tongue sole BAC library was constructed by an orderly mixing of the BAC clones. Using the specific primers designed to sox9 gene sequence, one positive single BAC clone was screened from the BAC library. In this study, the sox9-containing BAC clone, P17P21, was mapped to the long arms of a pair of homologous chromosomes of C. semilaevis using the technology of BAC-FISH. The result showed that there are two hybridization signals sites in a pair of autosomes of the female and male C.semilaevi, signals stable and specific. This study confirmed the effectiveness of the two-step-PCR screening system and achieved the positioning of sox9 gene on chromosomes of C. semilaevis. The results will facilitate subsequent in-depth studies on the sox9 gene of C. semilaevis and will provide an important basis for chromosomal identification of this species.