最新绿色荧光蛋白课件.ppt

1、 A topology diagram of the folding pattern in GFP.The-sheet strands are shown in light green,a-helices in blue,and connecting loops in yellow.The positions in the sequence that begin and end each major secondary structure element are also given.The anti-parallel strands(except for the interactions b

2、etween strands 1 and 6)make a tightly formed barrel.The green fluorescent protein GFP consists of 238 amino acids,linked together in a long chain.This chain folds up into the shape of a beer can.Inside the beer can structure the amino acids 65,66 and 67 form the chemical group that absorbs UV and bl

3、ue light,and fluoresces green.His48Gln94Arg96 Model of the fluorophore and its environment superposed on the MAD-phased electron density map at 2.2 resolution.The clear definition throughout the map allowed the chain to be traced and side chains to be well placed.The density for Ser65,Tyr66 and Gly6

4、7 is quite consistent with the dehydrotyrosine-imidazolidone structure proposed for the fluorophore.Many of the side chains adjacent to the fluorophore are labeled.475nm504nm 一种被修饰过的用来感应锌离子浓度的蓝色荧光蛋白：当红一种被修饰过的用来感应锌离子浓度的蓝色荧光蛋白：当红色的锌离子连接到蓝色的被修饰过的发色团上后，蛋白质会发色的锌离子连接到蓝色的被修饰过的发色团上后，蛋白质会发出亮度增强一倍的荧光从而形成容易被检测

5、到的可见信号。出亮度增强一倍的荧光从而形成容易被检测到的可见信号。Crystal structure of a Cu-bound green fluorescent protein Zn biosensor Crystal structure of a Zn-bound green fluorescent protein biosensor 88mM FIP-CBSM and 88 mM CaM(panels AC)88mM FIP-CBSM alone(panels DF).The photograph was taken 6 days post-inoculation using a DR

6、 Hand Lamp.The green fluorescent spots(against a background of red chlorophyll fluorescence)show the expanding foci of virus-infected cells.病毒介导的病毒介导的EGFP在烟草中的表达在烟草中的表达 RNA silencing of green fluorescent protein(GFP)(center)in leaves from Nicotiana benthamiana is suppressed by an animal(left;B2 prot

7、ein of flock house virus)or a plant(right)viral suppressor,leading to enhanced GFP expression(lighter green/yellow areas).GFP用于植物用于植物RNA沉默研究沉默研究 Moller 等研究发现等研究发现GFP 标记的标记的P.putidaRI 细菌主要细菌主要聚集在生物膜的表层，而聚集在生物膜的表层，而Acinetobacter sp.C6 则附着在则附着在生物膜底层生长。生物膜底层生长。C6:red R1:blueLocalisation of the ER target

8、ed form of GFP Roots from Arabidopsis plants transformed with a construct containing the cyclin B1;1promoter,cyclinB1;1 gene fused to GFP.Cells fluorescing in green are in mitosis 1997年年7月日本大阪大学的科研人员首次培育出月日本大阪大学的科研人员首次培育出能够夜里发光的含有绿色荧光蛋白的老鼠。能够夜里发光的含有绿色荧光蛋白的老鼠。1998年，年，Eduardo Kac通过转基因技术培育出第一条通过转基因技术培育

9、出第一条可以发出绿色荧光的狗。可以发出绿色荧光的狗。2004年年7月月12日韩国科学家成功培育出身体可发出荧光日韩国科学家成功培育出身体可发出荧光的转基因鸡。的转基因鸡。韩国科学家采用基因工程技术克隆的韩国科学家采用基因工程技术克隆的“荧光猫荧光猫”2007年登上年登上世界各大媒体的头条新闻。一旦暴露在紫外光线下，这种克隆猫世界各大媒体的头条新闻。一旦暴露在紫外光线下，这种克隆猫就能发红色的荧光就能发红色的荧光。2007年年1月月7日，东北农业大学刘忠华教授主持的转基因克隆猪课日，东北农业大学刘忠华教授主持的转基因克隆猪课题获得成功。中国首例绿色荧光蛋白转基因克隆猪成功产下题获得成功。中国首例

10、绿色荧光蛋白转基因克隆猪成功产下2头具有绿色头具有绿色荧光遗传特征的小猪。荧光遗传特征的小猪。The head of a barley plant transformed with a gene for green fluorescent protein(GFP)using the gene gun.(The red head is untransformed;the green head is transformed)基因枪介导的基因枪介导的GFP在小麦中的表达在小麦中的表达GFP canola(left)with a wild-type plant(right)under UV light.发绿光的油菜发绿光的油菜 Bacterial colony using various GFP and GFP-like proteins(from Tsien lab Web site).大脑彩虹图象抽象画大脑彩虹图象抽象画 此混杂又浓密的神经此混杂又浓密的神经元位于脑干的听觉区元位于脑干的听觉区 94 结束语结束语