scholarly journals Green Fluorescent Protein Tagging Drosophila Proteins at Their Native Genomic Loci With Small P Elements

Genetics ◽  
2004 ◽  
Vol 167 (4) ◽  
pp. 2143-2143
Author(s):  
PJ Clyne ◽  
J S Brotman ◽  
S T Sweeney ◽  
G Davis
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
P Elements ◽  
Green Fluorescent Protein Tagging ◽  
Green Fluorescent ◽  
Protein Tagging

scholarly journals Green Fluorescent Protein Tagging Drosophila Proteins at Their Native Genomic Loci With Small P Elements

Genetics ◽  
2003 ◽  
Vol 165 (3) ◽  
pp. 1433-1441
Author(s):  
Peter J Clyne ◽  
Jennie S Brotman ◽  
Sean T Sweeney ◽  
Graeme Davis
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Loss Of Function ◽  
P Elements ◽  
Native Proteins ◽  
A Genome ◽  
Potential Applications ◽  
Wide Scale ◽  
Green Fluorescent Protein Tagging ◽  
Green Fluorescent

Abstract We describe a technique to tag Drosophila proteins with GFP at their native genomic loci. This technique uses a new, small P transposable element (the Wee-P) that is composed primarily of the green fluorescent protein (GFP) sequence flanked by consensus splice acceptor and splice donor sequences. We demonstrate that insertion of the Wee-P can generate GFP fusions with native proteins. We further demonstrate that GFP-tagged proteins have correct subcellular localization and can be expressed at near-normal levels. We have used the Wee-P to tag genes with a wide variety of functions, including transmembrane proteins. A genetic analysis of 12 representative fusion lines demonstrates that loss-of-function phenotypes are not caused by the Wee-P insertion. This technology allows the generation of GFP-tagged reagents on a genome-wide scale with diverse potential applications.

scholarly journals Quantitative Imaging of B1 Cyclin Expression Across the Cell Cycle Using Green Fluorescent Protein Tagging and Epifluorescence

2020 ◽  
Vol 97 (10) ◽  
pp. 1066-1072
Author(s):  
Arnica Karuna ◽  
Francesco Masia ◽  
Sally Chappell ◽  
Rachel Errington ◽  
Andrew M. Hartley ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Quantitative Imaging ◽  
Green Fluorescent Protein Tagging ◽  
Green Fluorescent ◽  
Protein Tagging

scholarly journals Construction and Application of Epitope- and Green Fluorescent Protein-Tagging Integration Vectors for Bacillus subtilis

2002 ◽  
Vol 68 (5) ◽  
pp. 2624-2628 ◽  
Author(s):  
Marcus Kaltwasser ◽  
Thomas Wiegert ◽  
Wolfgang Schumann
Keyword(s):  
Bacillus Subtilis ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Pcr Amplification ◽  
Epitope Tag ◽  
Inner Membrane Protein ◽  
Green Fluorescent Protein Tagging ◽  
Green Fluorescent ◽  
Protein Tagging

ABSTRACT Here we describe the construction and application of six new tagging vectors allowing the fusion of two different types of tagging sequences, epitope and localization tags, to any Bacillus subtilis protein. These vectors are based on the backbone of pMUTIN2 and replace the lacZ gene with tagging sequences. Fusion of the tagging sequences occurs by PCR amplification of the 3′ terminal part of the gene of interest (about 300 bp), insertion into the tagging vector in such a way that a fusion protein will be synthesized upon integration of the whole vector via homologous recombination with the chromosomal gene. Three of these tagging sequences (FLAG, hemagglutinin, and c-Myc) allow the covalent addition of a short epitope tag and thereby detection of the fusion proteins in immunoblots, while three other tags (green fluorescent protein+, yellow fluorescent protein, and cyan fluorescent protein) are helpful in assigning proteins within one of the compartments of the cell. The versatility of these vectors was demonstrated by fusing these tags to the cytoplasmically located HtpG and the inner membrane protein FtsH.

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