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Cell Calcium ; 51(1): 57-64, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22133610

RESUMEN

While genetically encoded Ca(2+) indicators (GECIs) allow Ca(2+) imaging in model organisms, the gene expression is often under the control of a single promoter that may drive expression beyond, the cell types of interest. To enable more cell-type specific targeting, GECIs can be brought under the, control of the intersecting expression from two promoters. Here, we present the splitting and, reassembly of two representative GECIs (TN-XL and GCaMP2) mediated by the split intein from Nostoc, punctiforme (NpuDnaE). While the split TN-XL biosensor offered ratiometric Ca(2+) imaging, it had a, diminished Ca(2+) response relative to the native TN-XL biosensor. In contrast, the split GCaMP2, biosensor retained similar Ca(2+) response to the native GCaMP2. The split GCaMP2 biosensor was, further targeted to the pharyngeal muscles of Caenorhabditis elegans where Ca(2+) signals from feeding C. elegans, were imaged. Thus, we envision that increased cell-type targetability of GECIs is feasible with two, complementary promoters.


Asunto(s)
Calcio/metabolismo , Inteínas , Sondas Moleculares/metabolismo , Empalme de Proteína , Animales , Caenorhabditis elegans/metabolismo , Señalización del Calcio , Línea Celular , ADN Polimerasa III/metabolismo , Humanos , Indicadores y Reactivos , Mamíferos , Nostoc/metabolismo , Faringe/metabolismo
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