Making genes green: creating green fluorescent protein (GFP) fusions with blunt-end PCR products.
The jellyfish green fluorescent protein (GFP) has proven to be a useful tool in protein localization and trafficking studies. Fused to GFP, a protein of interest can be visualized and tracked in vivo through fluorescence microscopy. However, the process of making these fusion proteins is often tedious and painstaking. Here, we describe a simple and quick method for creating GFP fusion proteins using blunt-end PCR product ligation.
Department of Ophthalmology & Visual Science, Yale University School of Medicine, New Haven, CT 06520, USA.
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D001483:Base Sequence; D004274:DNA, Recombinant / Q000737:chemistry / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005057:Eukaryotic Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D005453:Fluorescence; D015870:Gene Expression / Q000235:genetics; D005796:Genes / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics
D004274:DNA, Recombinant; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C078383:endodeoxyribonuclease SrfI; D015252:Deoxyribonucleases, Type II Site-Specific