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半滑舌鳎含sox9基因BAC克隆的筛选及BAC-FISH定位
引用本文:孙建,陈松林,谢明树,邵长伟.半滑舌鳎含sox9基因BAC克隆的筛选及BAC-FISH定位[J].海洋水产研究,2014,35(1):34-40.
作者姓名:孙建  陈松林  谢明树  邵长伟
作者单位:[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
基金项目:国家自然科学基金重点项目(31130057)和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。

关 键 词:半滑舌鳎  sox基因  BAC文库  荧光原位杂交  染色体
收稿时间:2013/3/10 0:00:00
修稿时间:2013/4/9 0:00:00

Screening and BAC-FISH Locating of the sox9 Gene BAC Clone of Half-Smooth Tongue Sole Cynoglossus semilaevis
SUN Jian,',CHEN Song-lin,* XIE Ming-shu,SHAO Chang-wei.Screening and BAC-FISH Locating of the sox9 Gene BAC Clone of Half-Smooth Tongue Sole Cynoglossus semilaevis[J].Marine Fisheries Research,2014,35(1):34-40.
Authors:SUN Jian    CHEN Song-lin  * XIE Ming-shu  SHAO Chang-wei
Institution:2 ( College of Fisheries and I.ife Science, Dalian Ocean University, 116023) (z Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071)
Abstract: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.
Keywords:Cynoglossus semilaevis  sox9 gene  BAC library  Fluorescence in situ hybridization (FISH)  Chromosome
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