Designing and construction of a mammalian expression cassette containing monomeric green fluorescence protein gene in pCDNA3.1 by molecular cloning method

Abstract

Green fluorescence protein (GFP) is a small but a very stable protein which absorbs blue light and fluoresces green. It has a very negligible toxicity to the cell it expresses. Using these characteristics, protein tagging to the N or C terminus of GFP has revolutionized the research field. It has a wide range of application and various advantages in cell biological and proteomics study. Here in this dissertation we have successfully cloned the GFP gene within mammalian constitutive expression vector pcDNA3.1 which can be used to tag any protein of interest to the N or C terminus of GFP to visualize its localization and its interaction with other proteins through confocal microscope or any other fluorescence microscope.