REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
GFP-Booster Alexa Fluor 488 | Chromotek | Cat# gb2AF488-10 |
Bacterial and virus strains | ||
Sf9 cells | Expression systems | Cat#94-001F |
Subcloning Efficiency DH5α Competent Cells | Invitrogen | Cat#18265017 |
Chemicals, peptides, and recombinant proteins | ||
Cytochalasin-D | Sigma-Aldrich | Cat# C8273 |
1,6 Hexanediol (1,6-HD) | Sigma-Aldrich | Cat# 240117 |
RNase A | Roche | Cat# RNASEA-RO |
cOmplete Protease Inhibitor Cocktail, EDTA-free | Roche | Cat#5056489001 |
1X Dulbecco’s Phosphate Buffered Saline (PBS) solution without MgCl2 | Sigma-Aldrich | Cat# D8537 |
Potassium chloride | Merck | Cat#104935 |
DTT | Fermentas Life Sciences |
Cat#R0862 |
Benzonase | Produced in-house | N/A |
Tris | Carl Roth | Cat# 5429 |
Amylose resin | NEB | Cat# E8021S |
EDTA | Roche | Cat# 105063 |
Pipes | Applichem | Cat# A1079 |
monoGFP | Produced in-house | N/A |
Activation Buffer (AB) | York-Andersen et al., 2015 | N/A |
Schinder’s Drosophila medium | Gibco | Cat# 21720024 |
‘Wash Buffer A’ for Stellaris RNA FISH | LG Biosearch Technologies | Cat# SMF-WA1-60 |
‘Hybridisation Buffer’ for Stellaris RNA FISH | LG Biosearch Technologies | Cat# SMF-HB1-10 |
‘Probe Hybridisation Buffer’ for HCR V3.0 | Molecular Instruments | Bundled with custom probe set |
‘Probe Wash Buffer’ for HCR V3.0 | Molecular Instruments | Bundled with custom probe set |
‘Amplification Buffer’ for HCR V3.0 | Molecular Instruments | Bundled with custom probe set |
SlowFade Diamond Antifade Mountant with DAPI | Thermofisher Scientific | S36934 |
Protease inhibitor cocktail | Roche | Cat# CO-RO |
Amylose Resin | New England Biolabs | Cat# E8021S |
PreScission Protease GST 3C | GE Life Sciences | Cat# GE27-0843-01 |
Polyethene Glycol – 2000 | Merck | Cat#817018 |
Recombinant GFP-Me31B protein | This paper | N/A |
Recombinant GFP-Me31BΔN-ΔC protein | This paper | N/A |
Critical commercial assays | ||
Size exclusion chromatography using a HiLoad 16/600 Superdex 200 pg | GE Life Sciences | Cat# GE28-9893-35 |
Amicon Ultra-0.5 Centrifugal Filter Unit | Millipore | Cat# UFC5030 |
Deposited data | ||
Raw set of disordered regions from the Drosophila proteome | This paper | https://github.com/holehouse-lab/supportingdata/tree/master/2021/sankaranarayanan_me31b_2021 |
Experimental models: Organisms/strains | ||
D. melanogaster: y[1] w[∗]; P{w[+mC]=PTT-GB}me31B[CB05282] (Me31B::GFP) |
Bloomington Drosophila Stock Centre (Buszczak et al., 2007) | BDSC: 51530 FlyBase: FBst0051530 |
D. melanogaster: y,w, bcd-(ms2)6 (18), bcd-(ms2)6 (4); hsp83-MCP-RFP(4a) | Weil et al., 2006 | N/A |
D. melanogaster: GFP::Tral | Drosophila Genomics Resource Centre (Morin et al., 2001) | DGRC: 110584 Flytrap:G00089;DGRC:110584;RRID:DGGR_110584 |
D. melanogaster: y[1]; P{y[+mDint2] w[BR.E.BR]=SUPor-P} tral[KG08052] ry[506] / TM3, Sb[1] Ser[1] (tral1) |
Bloomington Drosophila Stock Centre (Wilhelm et al., 2005) | BDSC: 14933FlyBase: FBgn0041775 |
D. melanogaster: w[1118]; Df(3L)ED4483, P{w[+mW.Scer\FRT.hs3]=3’.RS5+3.3’}ED4483/ TM6C, cu[1] Sb[1] | Bloomington Drosophila Stock Centre (Wilhelm et al., 2005) | BDSC: 8070FlyBase: FBab0035731 |
Oligonucleotides | ||
Custom Stellaris FISH Probes for the 3’UTR of bcd RNA | Stellaris | See Table S2 (supplemental information). |
Molecular Instruments Custom HCR Probe for hb RNA | Molecular Instruments | DNA Custom kit Drosophila melanogasterlot number: 2690/B795GenBank: NM_169234.2, Alexa647, v3.0 kits |
Recombinant DNA | ||
Recombinant GFP-Me31B plasmid | This paper | N/A |
Recombinant GFP-Me31BΔN-ΔC plasmid | This paper | N/A |
Software and algorithms | ||
ImageJ | Schneider et al., 2012 | https://imagej.nih.gov/ij/ |
ImageJ Plugin Trackmate | Tinevez et al., 2017 | https://imagej.net/plugins/trackmate/ |
Rstudio/ R software | RStudio Team 2021 | http://www.rstudio.com/. |
HullRad | Fleming and Fleming, 2018 | N/A |
ABSINTH implicit solvent model | Vitalis and Pappu, 2009 | N/A |
CAMPARI Monte Carlo simulation (v3.0) | http://campari.sourceforge.net/V3/index.html | N/A |
SWISS-MODEL | Waterhouse et al., 2018 | N/A |
SOURSOP | https://soursop.readthedocs.io/ | N/A |
MDTraj | McGibbon et al., 2015 | N/A |
DSSP Algorithm | Kabsch and Sander, 1983 | N/A |
Protfasta | https://protfasta.readthedocs.io/ | N/A |
Metapredict | Emenecker et al., 2021 | N/A |
LocalCIDER | Holehouse et al., 2017 | N/A |
PIMMS simulation engine | Martin et al., 2020 | N/A |
Other | ||
Iberian recipe fly food | Produced in-house | N/A |
Oil 10 S (95 Series Halocarbon Oil) | VWR Chemicals | N/A |
Olympus FV3000 Confocal Laser Scanning Microscope | Olympus | FV3000 |
DeltaVision Core widefield microscope | Applied Precision, LLC | DeltaVision Core |
Custom built dual-trap optical tweezer instrument | Jahnel et al., 2011 | N/A |