生物技术通报2016，Vol.32Issue(3)：137-141,5.DOI:10.13560/j.cnki.biotech.bull.1985.2016.03.022
地高辛标记的黄鳝F64基因cRNA探针的制备及其应用
Preparation and Application of cRNA Probe Labeled with Digoxingenin for GeneF64 of Rice Field Eel
摘要
Abstract
The specific gene primers were designed based on the sequences of geneF64 of rice field eel,then the templates for synthesizing the cRNA probe were amplified,then recombinantF64/pGM-T plasmids were constructed and linearized by restriction enzymes, further the sense and anti-sense cRNA probes were synthesized byin vitro transcription with RNA polymerase,and finally labeled with Digoxigenin. The expression changes of gene F64during the gonadal development of rice field eel were detected byin situ hybridization. According to our results,the expression was only detected by the anti-senseF64 cRNA probe,and the earliest signal was observed in stage V of ovarian,but no more in early stage of ovarian formation. Our study indicated that the cRNA probes can be synthesized usingin vitro transcription,and the cRNA probes prepared in this study can be used for accurately detecting the spatial and temporal expression of geneF64.关键词
黄鳝F64基因/性腺发育/cRNA探针/原位杂交Key words
geneF64 of rice field eel/gonadal development/cRNA probe/in situhybridization引用本文复制引用
蒋骄云,田文斐,冯龙,曲宪成..地高辛标记的黄鳝F64基因cRNA探针的制备及其应用[J].生物技术通报,2016,32(3):137-141,5.基金项目
广西师范大学实验室管理创新与技术革新项目（2014GJ02），上海高校知识服务平台上海海洋大学水产动物遗传育种中心基金资助项目 （）
