Cognate putative nuclear localization signal effects strong nuclear localization of a GFP reporter and facilitates gene expression studies in Caenorhabditis elegans
Targeting a gene expression reporter, usually the green fluorescent protein (GFP), to the nucleus via a translationally fused nuclear localization signal (NLS) greatly facilitates recognition and identification of the reporter-expressing cells in Caenorhabditis elegans. Presently circulating nematod...
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Published in | BioTechniques Vol. 43; no. 5; pp. 596, 598, 560 - 600 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Natick, MA
Eaton
01.11.2007
Taylor & Francis Group |
Subjects | |
Online Access | Get full text |
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Abstract | Targeting a gene expression reporter, usually the green fluorescent protein (GFP), to the nucleus via a translationally fused nuclear localization signal (NLS) greatly facilitates recognition and identification of the reporter-expressing cells in Caenorhabditis elegans. Presently circulating nematode transcriptional gene expression vectors use the viral NLS from simian virus 40 (SV40) large T antigen. This NLS, however fails to ensure sufficient localization of the GFP peptide to the nucleus. We modified the common transcriptional reporter SV40 NLS-GFP by adding to its C terminus a cognate putative NLS from the transcription factor egl-13. The EGL-13 NLS effected clear contrast in fluorescence intensity between the nucleus and the cytoplasm in cells with strong reporter signal and efficiently highlighted the nucleus in tissues with weak reporter expression in a wide range of tested tissues. The SV40 NLS-GFP-EGL-13 NLS vector should become a valuable tool for gene expression studies in C. elegans. |
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AbstractList | Targeting a gene expression reporter, usually the green fluorescent protein (GFP), to the nucleus via a translationally fused nuclear localization signal (NLS) greatly facilitates recognition and identification of the reporter-expressing cells in Caenorhabditis elegans. Presently circulating nematode transcriptional gene expression vectors use the viral NLS from simian virus 40 (SV40) large T antigen. This NLS, however fails to ensure sufficient localization of the GFP peptide to the nucleus. We modified the common transcriptional reporter SV40 NLS-GFP by adding to its C terminus a cognate putative NLS from the transcription factor egl-13. The EGL-13 NLS effected clear contrast in fluorescence intensity between the nucleus and the cytoplasm in cells with strong reporter signal and efficiently highlighted the nucleus in tissues with weak reporter expression in a wide range of tested tissues. The SV40 NLS-GFP-EGL-13 NLS vector should become a valuable tool for gene expression studies in C. elegans. |
Author | SCHLEGEL, Robert A HANNA-ROSE, Wendy LYSSENKO, Nicholas N |
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Cites_doi | 10.1242/dev.126.1.169 10.1074/jbc.M601718200 10.1093/genetics/157.3.1217 10.1016/j.cell.2004.09.001 10.1016/S1388-1981(02)00324-4 10.1146/annurev.biochem.67.1.509 10.1016/0092-8674(87)90500-9 10.1104/pp.007971 10.1016/j.cub.2007.05.024 10.1895/wormbook.1.45.1 |
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Keywords | Caenorhabditis elegans Signal Reporter gene Helmintha Nemathelminthia Invertebrata Gene expression Localization Nematoda |
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References | e_1_3_4_4_1 e_1_3_4_3_1 Fire A. (e_1_3_4_6_1) 1998 e_1_3_4_9_1 e_1_3_4_8_1 e_1_3_4_7_1 e_1_3_4_5_1 e_1_3_4_12_1 e_1_3_4_13_1 e_1_3_4_10_1 e_1_3_4_11_1 Hobert O. (e_1_3_4_2_1) 2006 Bargmann C.I. (e_1_3_4_14_1) 1995 |
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SubjectTerms | Animals Biological and medical sciences Caenorhabditis elegans - genetics Cell Nucleus - metabolism Diverse techniques Fundamental and applied biological sciences. Psychology Gene Expression Genes, Reporter Green Fluorescent Proteins - metabolism Molecular and cellular biology Nuclear Localization Signals - metabolism Protein Transport |
Title | Cognate putative nuclear localization signal effects strong nuclear localization of a GFP reporter and facilitates gene expression studies in Caenorhabditis elegans |
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