Retroviral expression of green fluorescent protein.

摘要: Publisher Summary Retroviral vectors are powerful tools for gene transfer in vitro and vivo. They have a number of convenient features that explain their multiple experimental clinical applications. Most importantly, retrovirus-mediated leads to stable proviral integration the target cell genome, allowing transmission foreign genetic material progeny infected cells. can be modified by deleting all viral protein-coding sequences (gag, pol, env) without significant decrease efficiency infection. Helper-free virus stocks obtained safe using retrovirus packaging lines, which supply proteins required vector transmission. In addition, defective recombinant retroviruses allow efficient into broad range mammalian avian cells derived from different species tissues. It is not surprising discovery green fluorescent protein (GFP) its variants had an immediate impact on field retrovectorology. Detection GFP performed living samples, amenable real-time analysis molecular events. When expressed either eukaryotic or prokaryotic illuminated blue ultraviolet (UV) light, emits bright signal easily detected fluorescence microscopy, flow cytometry, other imaging techniques. Light-stimulated independent does require additional cofactors, substrates, products. Therefore, superior markers used selection viable such as antibiotic resistance (e.g., neo, hyg), requires extensive culture selective medium, surface markers, staining with specific antibodies.