摘要
本研究通过转录法合成了4个发育基因eve、ben、Dfd和Scr的cRNA探针,并对蜜蜂幼虫进行了原位杂交。结果显示:eve、ben、Dfd和Scr基因在幼虫中都有表达,eve与ben基因是在体节上呈横竖条纹状表达而dfd与Scr基因是在体节两侧边缘呈点状表达。结果表明这4个基因在蜜蜂幼虫阶段也参与了体节的发育和形态建成。基于此,还对胚胎整体原位杂交中的一些技术细节,如探针标记、探针保存、信号检测、固定时间、消化时间和操作手法等进行了讨论。
Four cRNA probes including the pair-rule genes such as eve, the segmental polarity genes such as ben and the homeotic genes such as dfd or scr were synthetized through a transcriptional method and then were hybridized in situ with the transcripts in honeybee larvae. The results showed that genes eve, ben, dfd and scr had an obvious expression in honeybee at larval stage, respectively. Gene eve or gene ben had a striped expression on segments horizontally and vertically while gene dfd or gene scr had a dotted expression on bilateral boundary of segments. The results indicated that genes such as eve, ben, Dfd and Scr were also involved in segmental development and morphological establishment of honeybee at larval stage. Some technical details about whole mount embryo in situ hybridization were also discussed, for example, probe labelling, probe preservation, signal detection, time length for sample fixation by paraformalclehyde, time length for sample digestion by proteinase, and operation technique.
出处
《中国蜂业》
2014年第Z2期4-10,共7页
Apiculture of China
基金
福建省自然科学基金资助项目(2009N0003)
关键词
蜜蜂
幼虫
发育基因
原位杂交
Honey bees
Larva
Developmental gene
Hybridization in situ
