线虫荧光标记稳定表达系的建立

Establishment of a stable C.elegans stain expressing PHluorin labeled vesicle

目的:建立稳定表达的PHluorin标记的线虫种系,为囊泡在线虫ALA神经元上分泌机制的研究提供模型。方法:采用了国际先进的线虫转基因技术,将构建的Pida-1IDA-1:PHluorin质粒通过显微注射到线虫的母代,通过筛选后得到稳定表达的种系。结果:通过DIC显微镜整体检测和全内反射荧光成像技术(Tirfm)细胞检测,蛋白表达的位置正确,通过高倍数体式显微镜确定稳定种系中阳性率高达99%。结论:建立了一个稳定表达的荧光标记线虫种系,为进一步在线虫上研究囊泡分泌提供了很好的模型。

Objective：To establish a stable C.elegans stain expressing PHiuorin labeled vesicles and to lay a foundation for the investigating the vesicles release mechanism in the ALA neuron of C.elegans.Methods：Using the microinjection,the Pida-1 IDA-1：PHluorin was microinjected to the c.elegans,and positive strains were screened.Results：The DIC microscope and Tiffin detection showed that over 99% C.elegans was expressed the marked protein,and further distribution results showed that most of the marked protein was stored in the ALA neuron vesicles.Conclusion：A stable strain expressing the phluorin was established,and provided a good model for studying vesicles release mechanism in the C.elegans.