Journal: Nature methods
Article Title: A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum
Figure Lengend Snippet: Fluorescence imaging of mNeonGreen (mNG) fusion vectors. C-terminal mNG fusion constructs (with respect to the fluorescent protein); for each fusion, the linker amino acid length is indicated after the name of the targeted organelle or fusion partner: ( a ) mNG-Annexin (A4)-C-12 (human; plasma membrane); ( b ) mNG-β-actin-C-18 (human; actin cytoskeleton); ( c ) mNG-β-Catenin-C-20 (mouse; tight junctions); ( d ) mNG-CAAX-C-5 (20-amino acid farnesylation signal from c-Ha-Ras; plasma membrane); (e) mNG-CAF1-C-10 (mouse chromatin assembly factor); ( f ) mNG-Caveolin1-C-10 (human); ( g ) mNG-Endosomes-C-14 (human RhoB GTPase); ( h ) mNG-Fascin-C-10 (human; actin bundling); ( i ) mNG-Fibrillarin-C-7 (human; nucleoli); ( j ) mNG-FilaminA-C-14 (human; cytoskeleton); ( k ) mNG-LAMP1-C-20 (rat; lysosomal membrane glycoprotein 1; lysosomes); ( l ) mNG-Clathrin-C-15 (human, light chain B); ( m ) mNG-Myotilin-C-14 (human; actin filaments); ( n ) mNG-PCNA-C-19 (human; replication foci); ( o ) mNG-Plastin-C-10 (human; actin binding); ( p ) mNG-Rab4a-C-7 (human; endosomes); ( q ) mNG-LC3B-C-7 (rat light chain; autophagosoms; ( r ) mNG-Talin-C-18 (mouse; focal adhesions); ( s ) mNG-Tubulin-C-35 (human; microtubules); ( t ) mNG-ZO1-C-14 (human; tight junctions). The cell line used for expression of C-terminal mNG constructs was Madin-Darby canine kidney (MDCK; ATCC, CCL-34) cells in panels ( c ) and ( t ). HeLa CCL2 (ATCC) cells were used in the remaining panels. Scale bars represent 10 μm.
Article Snippet: Thus, to prepare mNeonGreen C-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: annexin A4 (12), NheI and BspEI (Alen Piljic, EMBL, Heidelberg, Germany; NM_001153.3); β-actin (7), NheI and BglII (human β-actin cDNA source: Clontech; NM_001101.3); β-catenin (20), XhoI and BamHI (mouse β-catenin cDNA source: Origene, Rockville, MD; NM_001165902.1); 20 amino acid farnesylation signal from c-Ha-Ras (CAAX; 5), AgeI and BspEI (c-Ha-Ras cDNA source: Clontech; NM_001130442.1); CAF1 (10), AgeI and BspEI (mouse chromatin assembly factor cDNA source: Akash Gunjan, Florida State University; NM_013733.3); caveolin 1 (10), NheI and BglII (human caveolin 1 cDNA source: Origene; NM_001753); endosomes (14), NheI and BspEI (endosomes cDNA source: Clontech; NM_004040.2); fascin (10), BspEI and BamHI (human fascin cDNA source: Origene; NM_003088.2); fibrillarin (7), AgeI and BspEI (fibrillarin cDNA source: Evrogen, Moscow, Russia; NM_001436.3); filamin A (14), BspEI and HindIII (human filamin cDNA source: David Calderwood, Yale University; NM_001456.3); human lysosomal membrane glycoprotein 1 (20), BamHI and NotI (LAMP1; George Patterson, NIH, Bethesda MD, U.S.A.; NM_012857.1); human light chain clathrin (15), NheI and BglII (human clathrin light chain cDNA source: George Patterson, NIH; NM_001834.2); human myotilin, AgeI and BspEI (MYOT; Origene; NM_006790.1); PCNA (19), AgeI and BspEI (proliferating cell nuclear antigen cDNA source: David Gilbert, FSU; NM_002592.2); plastin (10), BspEI and XhoI (human plastin 1 (fimbrin) cDNA source: Origene; NM_002670.1); canine Rab4a, BglII and BamHI (Rab4a cDNA source: Viki Allen, U. Manchester, UK; NM_004578.2); LC3B (7), AgeI and BspEI (rat LC3B cDNA source: Jenny M. Tam, Harvard University; U05784.1); talin (22) AgeI and BspEI (mouse talin 1 cDNA source: Clare Waterman, NIH; NM_011602.5); α-tubulin (18), NheI and BglII (human α-tubulin cDNA source: Clontech; NM_006082).
Techniques: Fluorescence, Imaging, Construct, Binding Assay, Expressing