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פותח על ידי קלירמאש פתרונות בע"מ -
A genetic system for detection of protein nuclear import and export
Year:
2000
Source of publication :
Nature Biotechnology
Authors :
גפני, ידידיה
;
.
Volume :
18
Co-Authors:
Rhee, Y., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gurel, F., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gafni, Y., Department of Genetics, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Dingwall, C., Department of Pharmacology, State University of New York, Stony Brook, NY 11794-8651, United States, Neurosciences Research Department, SmithKline-Beecham Pharmaceutical, Harlow, Essex, United Kingdom
Citovsky, V., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Facilitators :
From page:
433
To page:
437
(
Total pages:
5
)
Abstract:
We have developed a simple genetic assay to detect active nuclear localization (NLS) and export signals (NES) on the basis of their function within yeast cells. The bacterial LexA protein was modified (mLexA) to abolish its intrinsic NLS and fused to the activation domain of the yeast Gal4p (Gal4AD) with or without the SV40 large T-antigen NLS. In the import assay, if a tested protein fused to mLexA-Gal4AD contains a functional NLS, it will enter the cell nucleus and activate the reporter gene expression. In the export assay, if a tested protein fused to mLexA-SV40 NLS-Gal4AD contains a functional NES, it will exit into the cytoplasm, decreasing the reporter gene expression. We tested this system with known NLS and NES and then used it to demonstrate a NES activity of the capsid protein of a plant geminivirus. This approach may help to identify, analyze, and select for proteins containing functional NLS and NES.
Note:
Related Files :
Agrobacterium
Miridae
nuclear localization signal
Saccharomyces cerevisiae Proteins
Tomato yellow leaf curl virus
עוד תגיות
תוכן קשור
More details
DOI :
10.1038/74500
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
25529
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:15
Scientific Publication
A genetic system for detection of protein nuclear import and export
18
Rhee, Y., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gurel, F., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gafni, Y., Department of Genetics, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Dingwall, C., Department of Pharmacology, State University of New York, Stony Brook, NY 11794-8651, United States, Neurosciences Research Department, SmithKline-Beecham Pharmaceutical, Harlow, Essex, United Kingdom
Citovsky, V., Dept. of Biochem. and Cell Biology, Inst. for Cell and Devmtl. Biology, State University of New York, Stony Brook, NY 11794-5215, United States
A genetic system for detection of protein nuclear import and export
We have developed a simple genetic assay to detect active nuclear localization (NLS) and export signals (NES) on the basis of their function within yeast cells. The bacterial LexA protein was modified (mLexA) to abolish its intrinsic NLS and fused to the activation domain of the yeast Gal4p (Gal4AD) with or without the SV40 large T-antigen NLS. In the import assay, if a tested protein fused to mLexA-Gal4AD contains a functional NLS, it will enter the cell nucleus and activate the reporter gene expression. In the export assay, if a tested protein fused to mLexA-SV40 NLS-Gal4AD contains a functional NES, it will exit into the cytoplasm, decreasing the reporter gene expression. We tested this system with known NLS and NES and then used it to demonstrate a NES activity of the capsid protein of a plant geminivirus. This approach may help to identify, analyze, and select for proteins containing functional NLS and NES.
Scientific Publication
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