线虫荧光标记稳定表达系的建立

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。

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

Objective:To establish a stable C.elegans stain expressing PHiuorin labeled vesicles and to lay a foundation for the in- vestigating the vesicles release mechanism in the ALA neuron of C.elegans.Methods:Using the microinjection,the Pida-1 IDA-1:PHluo- rin 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.