Functional expression of green fluorescent protein derivatives in Halobacterium salinarum.
We investigated the applicability of the green fluorescent protein (GFP) of Aequorea victoria as a reporter for gene expression in an extremely halophilic organism: Halobacterium salinarum. Two recombinant GFPs were fused with bacteriorhodopsin, a typical membrane protein of H. salinarum. These fusion proteins preserved the intrinsic functions of each component, bacteriorhodopsin and GFP, were expressed in H. salinarum under conditions with an extremely high salt concentration, and were proved to be properly localized in its plasma membrane. These results suggest that GFP could be used as a versatile reporter of gene expression in H. salinarum for investigations of various halophilic membrane proteins, such as sensory rhodopsin or phoborhodopsin.
Nomura
S
S
Biomolecular Engineering Research Institute, Osaka, Japan.
Harada
Y
Y
D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000235:genetics* / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D019766:Genes, Archaeal; D049452:Green Fluorescent Proteins; D006217:Halobacterium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D019843:Archaeal Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012965:Sodium Chloride; D001436:Bacteriorhodopsins