摘要
为研究doublesex(dsx)基因在不同生殖状态的蚤状溞的时空表达状况,提取不同生殖状态的蚤状溞RNA,采用RT-PCR和Quantitative real-time PCR检测了dsx在不同生殖状态溞体的表达差异,并通过体外转录制备DIG标记的Doublesex的RNA探针;采用整体原位杂交方法研究dsx基因在不同生殖状态的蚤状溞的时空表达状况。结果显示,RT-PCR检测发现dsx在雌雄溞体没有性别特异性的选择性拼接,只有一种相同的转录产物。Real-time PCR显示,dsx在雄溞中表达水平最高,是孤雌溞的2.6倍,两性雌溞的4.9倍,三者之间有显著性差异,表现出性别差异性表达。整体原位杂交发现,在所取样的不同生殖状态溞体中dsx基因也均有表达,但其部位与表达量不同,且在雌雄溞体中表现出性别差异。在雄溞的第一触角和第一胸肢显现得尤为明显,且在复眼中也有表达。孤雌溞和两性雌溞对应部位表达则相对较弱。结果表明,dsx基因很有可能在调控蚤状溞生殖转换和性别分化上起到很大作用。
To analyze the expression pattern of Doublesex （dsx） of Daphnia pulex in different reproductive modes, we extracted total RNA of different reproductive modes＇ Daphnia pulex. Using RT-PCR and Quantitative real-time PCR analyzed dsx expression in different reproductive modes＇ daphnias. In addition, we synthesized DIG labeled RNA probe of dsx by in vitro transcription and used whole mount in situ hybridization researching dsx location expression in different reproductive modes＇ daphnia. Results showed that dsx in male and female had no gender specificity of alternative splicing, only a same transcription. Realtime PCR showed dsx highest expression level was in male, which was parthenogenetic female＇ s 2.6 times and 4.9 times that of the female. They had significant difference （P 〈 0. 05） and gender differences in expression each other. Whole mount in situ hybridization found that dsx gene was expressed in the somatic tissure in different from reproductive modes＇ daphnia, but the difference in quantity and parts. The expression also showed the gender differences in male and female. In male the signals were revealed in the first antenna, the first thoracic limb and the compound eye. The parthenogenetic female and sexual female corresponding sites＇ expressions were relatively weak. These suggested that Doublesex gene likely played a significant role in alternation of generation and sexual differentiation.
出处
《水产学报》
CAS
CSCD
北大核心
2013年第11期1655-1662,共8页
Journal of Fisheries of China
基金
国家自然科学基金项目(31172043)
浙江省自然科学基金项目(LY12C19003)
上海市科技创新行动计划项目(12391900700)
公益性行业(农业)科研专项(200903056)
宁波市科技局重大择优项目(2012C10020)
