基于激光共聚焦显微镜的绿色荧光融合蛋白质微米环形貌分析  

Analysis of Micro Ring Shape of Green Fluorescent Fusion Protein Based on Laser Confocal Microscope

作　　者：胡坤[1] HU Kun(Scientific Research Center of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院科研中心,安徽蚌埠233000

出　　处：《通化师范学院学报》2020年第12期45-51,共7页Journal of Tonghua Normal University

基　　金：蚌埠医学院自然科学基金项目(BYKY1761).

摘　　要：目的:分析绿色荧光融合蛋白质微米环形貌.方法:选取绿色荧光蛋白质试剂,构建融合蛋白质原核表达载体,配制凝胶,对融合蛋白质进行纯化,加强融合蛋白质荧光效应,使其达到微米环表达条件,对融合蛋白质进行酯化反应,利用激光共聚焦显微镜表征微米环形貌,计算单峰状态和聚集状态的分子量.结果:绿色荧光蛋白质融合后,微米环单峰状态和聚集状态的分子量均符合产物电泳的理论值.结论:融合蛋白质微米环形貌规整,具有较强荧光,宽度一直保持30 nm左右,微米环配体分子两端分别为生物素和罗丹明B分子,中间为3个重复单元的配体Rh4Bio,连接多肽链具有柔性,以纳米线状结构的二聚体形式存在.Objective:to analyze the micromorphology of color fluorescence fusion protein.Methods:green fluorescent protein reagent was used to construct the fusion protein prokaryotic expression vector,and the gel was configured to purify the fusion protein,enhance the fluorescence effect of the fusion protein,make it reach the micron ring expression condition,esterify the fusion protein,characterize the micron ring appearance by the light confocal microscope,and calculate the molecular weight of the single peak state and the aggregation state.Results:after fusion,the molecular weights of the single peak state and ag⁃gregation state of the green fluorescent protein conformed to the theoretical value of electrophoresis.Conclusion:the fusion protein has regular micro ring appearance,strong fluorescence,and the width is about 30 nm.The ends of the ligand are biotin and Rhodamine B,and the middle is the ligand Rh4Bio with three repeat units.The peptide chain is flexible and exists in the form of dimer with nanowire structure.

关 键 词：激光共聚焦显微镜 绿色荧光蛋白质 蛋白质微米环 形貌分析 荧光探针 

分 类 号：TH742[机械工程—光学工程]