细胞生物学：第三章 细胞生物学的研究方法.pptVIP

Adv Rem was used to infect Hep G2 cells Expression of H2B-GFP before and after UV-B-induced apoptosis. A431 cells were stably transfected with H2B-GFP fusion protein (K/H2B-GFP). H2B-GFP is expressed only within the nuclei. In the right panel an apoptotic K/H2B-GFP cell is shown after UV-B irradiation. The “Blebs and bodies forming” phase is shown. Noticeably, redistribution of SLE self antigens are among the remarkable changes that occur during apoptosis. Images were collected in real time by laser confocal microscopy. GFP expression in haematopoietic colonies from transgenic mice carrying the green fluorescent protein (GFP)-reporter gene under the control of Kit (stem cell factor receptor) regulatory elements. These mouse lines provide useful means to mark rare adult multipotential cells which, upon transplantation, can be monitored for their ability to engraft, differentiate and regenerate different tissues. （三）荧光共振能量转移可实时观察细胞内蛋白与蛋白的相互作用 荧光共振能量转移（fluorescence resonance energy transfer, FRET) Cy3-cy5, Alexa546-Alexa647; CFP-YEP, BFP-EGFP B.荧光共振能量转移可实时观察细胞内蛋白与蛋白的相互作用 荧光共振能量转移（florescence resonance energy transfer, FRET) Cy3-cy5, Alexa546-Alexa647; CFP-YEP, BFP-EGFP Excitation: 402nm emission 475nm Excitation: λmax＝395nm和λmin＝470nm Emission:λ＝509nm B.荧光共振能量转移可实时观察细胞内蛋白与蛋白的相互作用 荧光共振能量转移（florescence resonance energy transfer, FRET) Cy3-cy5, Alexa546-Alexa647; CFP-YEP, BFP-EGFP Excitation: 402nm emission 475nm Excitation: λmax＝395nm和λmin＝470nm Emission:λ＝509nm 第五节 细胞功能基因组学研究技术 一. DNA基本研究技术 （一）限制性核酸内切酶可特定位点切断DNA分子 （常规的分子生物学技术之一） （二）不同支持物的电泳分离不同 大小的DNA分子 （常规的分子生物学技术之二） （三）聚合酶链式反应对特定的DNA序列进行复制 聚合酶链式反应（polymerase chain reaction, PCR) DNA RNA mRNA tRNA rRNA Protein Replication Transcription Translation Reversed transcription DNA in vitro amplification (PCR) The discovery of Taq enzyme. Taq DNA polymerase is a thermostable DNA polymerase that prossesses a 5’- 3’polymerase activity and a Double strand specific 5’-3’ exonuclease activity. 94C