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

作　　者：孙建[1,2] 陈松林[2] 谢明树[2] 邵长伟[2] 

机构地区：[1]大连海洋大学水产与生命学院,116023 [2]农业部海洋渔业可持续发展重点实验室中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《渔业科学进展》2014年第1期34-40,共7页Progress in Fishery Sciences

基　　金：国家自然科学基金重点项目(31130057);国家自然科学基金项目(41006107)共同资助

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两个杂交信号位点,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,其结果为揭示sox9基因参与鱼类性别控制的机制提供了重要基础。A two-step PCR screening system of the half-smooth tongue sole Cynoglossus semilaevis BAC library was constructed by an orderly mixing of the BAC clones. Using the spe- cific primers designed to sox9 gene sequence, one positive single BAC clone was screened out 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 signal sites in a pair of auto- somes of the female and male C. semilaevi, and the signals are 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 further studies on the sox9 gene of C. semilaevis and will provide an important basis for chromosomal identification of this species.

关 键 词：半滑舌鳎 SOX9基因 BAC文库 荧光原位杂交 染色体 

分 类 号：Q959.486[生物学—动物学]