Tubulin gal4

Manufactured by Bloomington Drosophila Stock Center

Tubulin-Gal4 is a genetic tool used in Drosophila (fruit fly) research. It is a Gal4 driver line that expresses the Gal4 transcription factor under the control of the tubulin promoter, which drives ubiquitous expression throughout the fly. The Tubulin-Gal4 line can be used to drive the expression of any gene of interest downstream of the UAS (Upstream Activating Sequence) promoter in a wide range of Drosophila tissues and cell types.

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Maternal-tubulin-Gal4 (2 citations) Tubulin-GAL4 driver (2 citations)

14 protocols using tubulin gal4

Drosophila Tau Transgenic Lines

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GMR-GAL4, elav-GAL4, actin-GAL4, tubulin-Gal4, tau^HM05101, tubP-GAL80^ts, UAS-Dcr-2, tau^Df(3R)MR22, Df(3R)BSC499, and UAS-hTau23 (UAS-MAPT.A59A) were obtained from the Bloomington Stock Center. UAS-dTau::GFP and tau^EP3597 were a gift of D. St. Johnston (University of Cambridge, Great Britain). Appl-Gal4 was obtained from Laura Torroja (Universidad Autonoma de Madrid, Spain). tau^GD25023 was received from the Vienna Drosophila RNAi Center (Vienna, Austria). olk¹ mutants are described in (Bettencourt da Cruz et al., 2005 (link)). Flies were raised under standard conditions at 25°C unless specifically described. To create the MAP60 and MAP205-expressing flies, we used LD02709 for MAP60 and LD12965, which were cloned into pUAST. Transgenic flies were generated by P-element transformation using Best Gene Inc. (Chino Hills, CA).

Bolkan B.J, & Kretzschmar D. (2014). Loss of Tau results in defects in photoreceptor development and progressive neuronal degeneration in Drosophila. Developmental neurobiology, 74(12), 1210-1225.

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Drosophila Genetic Crosses for Functional Analysis

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The following lines were acquired from the Bloomington Stock Center: w¹¹¹⁸ (BL# 38690), hsFlp,hsIscel/CyO (BL # 6934), nanos-Gal4 (BL# 32179), Tubulin-Gal4 (BL# 5138), eyeless-Gal4 (BL# 8227), In(1)white^m4 (BL# 807), Sb[V] (BL# 878), Cre (BL# 1501) lid¹⁰⁴²⁴ (BL# 12367), lid^k06801 (BL# 10403). PSR^FM1 was provided by Kristin White (Massachusetts General Hospital, Charlestown, MA), FRT40A, UTX^{1 (link)} was provided by Andreas Bergmann (M. D. Anderson Cancer Center, Houston, TX), FRT40A, UTX^Δ was provided by Jürg Müller (MPI of Biochemistry, Chromatin and Chromosome Biology, Martinsried, Germany).

Shalaby N.A., Sayed R., Zhang Q., Scoggin S., Eliazer S., Rothenfluh A, & Buszczak M. (2017). Systematic discovery of genetic modulation by Jumonji histone demethylases in Drosophila. Scientific Reports, 7, 5240.

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Live Imaging of Drosophila Cell Junctions and Dynamics

Cited 7 times

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The following markers were used for live imaging: endo-DE-cadherin:GFP (Huang et al., 2009 (link)), ubi-DE-cadherin:GFP (Oda and Tsukita, 2001 (link)), UAS-DE-cadherin:GFP (gift of N. Gorfinkiel, Consejo Superior de Investigaciones Cientificas–Universidad Autónoma de Madrid, Madrid, Spain), UAS-p120-catenin:GFP (Bloomington Drosophila Stock Center; Myster et al., 2003 (link)), UAS-actin:RFP (Simões et al., 2006 (link); gift of N. Gorfinkiel), sqh-GFP:moesin (Kiehart et al., 2000 (link)), UAS-mCherry:moesin (Millard and Martin, 2008 (link)), sqh-sqh:GFP (Royou et al., 2004 (link)), sqh-sqh:mCherry (Martin et al., 2009 (link)), sqh-GFP:rok^K116A (Simões et al., 2010 (link)), UAS-GFP:clc (Chang et al., 2002 (link)), 20XUAS-shi^ts1:GFP (Pfeiffer et al., 2012 (link)), UAS-Apoliner (Bloomington Drosophila Stock Center; Bardet et al., 2008 (link)), UAS-GFP (gift of U. Tepass, University of Toronto, Toronto, Canada), 10XUAS-myr:GFP (Bloomington Drosophila Stock Center; Pfeiffer et al., 2010 (link)), UAS-GCaMP3 (Bloomington Drosophila Stock Center; Tian et al., 2009 (link)), and UAS-arf6:GFP (this study). daughterless-Gal4 or tubulin-Gal4 (Bloomington Drosophila Stock Center) were used to drive ubiquitous expression of all UAS transgenes. tubulin-Gal4 was used for all overexpression experiments. yellow white flies were used for controls.

Hunter M.V., Lee D.M., Harris T.J, & Fernandez-Gonzalez R. (2015). Polarized E-cadherin endocytosis directs actomyosin remodeling during embryonic wound repair. The Journal of Cell Biology, 210(5), 801-816.

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Generating Drosophila Genetic Reporters

Cited 1 time

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WT flies used were Canton S (CS). Lar and sns mutants have been previously described: Lar^13.2, Lar⁴⁵¹, and sns^xb3 (gift of Dr. Susan Abmayr). The following lines were obtained from the Bloomington Stock Center: sns^Df, C155-GAL4, tubulin-GAL4, OK107-GAL4, sns^{MiMIC MI03001}, Lar^MiMIC02154, sns^EY08142, UAS-Lar RNAi (TRiP.HMS02186), UAS-Lar RNAi (TRiP.HMS00822), and UAS-Kirre RNAi (TRiP.HMC05791). Sns RNAi lines were from the Vienna Drosophila Resource Center: UAS-Sns RNAi (KK109442) and UAS-Sns RNAi (GD877). T2A-GAL4 lines were generated as described in Diao et al., 2015 (link). Briefly, flies carrying the MiMIC insertion were crossed with flies bearing the triplet ‘Trojan exon’ donor. The F₁ males from this cross carrying both genetic components were crossed to females carrying germline transgenic sources of Cre and ϕC31. The F₂ males from this cross that had all four genetic components were then crossed to a UAS-2xEGFP reporter line, and the resulting progeny were screened for T2A-GAL4 transformants.

Bali N., Lee H.K, & Zinn K. (2022). Sticks and Stones, a conserved cell surface ligand for the Type IIa RPTP Lar, regulates neural circuit wiring in Drosophila. eLife, 11, e71469.

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Fly Genetic Manipulation Protocols

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The UAS-Datlastin-myc and UAS-BiP-sfGFP-HDEL fly lines used in this study were described previously (Orso et al., 2009 (link); Summerville et al., 2016 (link)). The Gal4 strains used were: tubulin-Gal4, elav-Gal4, D42-Gal4, and GMR-Gal4 obtained from Bloomington Drosophila Stock Center. To increase protein expression, experimental crosses were performed at 28°C, except for crosses with GMR-Gal4 which were performed at 25°C. Control genotypes included promoter-Gal4/+ individuals. Fly food was prepared using NUTRI-fly-IF mixture (Genesee Scientific), according to the manufacturer instructions.

Montagna A., Vajente N., Pendin D, & Daga A. (2020). In vivo Analysis of CRISPR/Cas9 Induced Atlastin Pathological Mutations in Drosophila. Frontiers in Neuroscience, 14, 547746.

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Drosophila RNAi and Heteroplasmic Phenotypes

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All stocks were maintained at 25 °C and at 70% relative humidity in 12-h light/dark cycles and were fed standard yeast-molasses-agar medium. The following fly lines were obtained from Bloomington Drosophila Stock Center: UAS-park^RNAi (Bloomington stock #: 31259), UAS-rok^RNAi (Bloomington stock #: 34324), TH-GAL4 (Bloomington stock #: 8848), tubulin-GAL4 (Bloomington stock #5138). Canton (S) and heteroplasmic mtColI^T300I flies were generously provided by Dr. Thomas Hurd (University of Toronto).

Moskal N., Riccio V., Bashkurov M., Taddese R., Datti A., Lewis P.N, & Angus McQuibban G. (2020). ROCK inhibitors upregulate the neuroprotective Parkin-mediated mitophagy pathway. Nature Communications, 11, 88.

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Drosophila Transgenesis and Lifespan Assay

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Fly culture and transgenesis were performed using standard procedures. Rtnl1-PB cDNA was cloned in the pUAST vector for Drosophila transgenesis in frame with a HA tag.
Primers used:
Rtnl1-PB HA 5′-AGCTGAATTCATGTACCCATACGATGTTCCTGACTATGCGGGCTCCGCATTTGGTGAAACC-3′
Rtnl1-PB 5′-AGCTTCTAGATTACTTGTCCTTCTCAGAC-3′
Several transgenic lines for UAS-HA-Rtnl1 were generated and tested. Drosophila strains GMR-Gal4, D42-Gal4, tubulin-Gal4, arm-Gal4, pUASp:Lys-GFP-KDEL were obtained from the Bloomington Drosophila Stock Center. UAS-Rtnl1-RNAi lines were obtained from the Vienna Drosophila RNAi Center (v7866 and v33919). Lifespan experiments were performed with 200 animals for each genotype. Flies were collected 1 day after eclosion and placed in vials containing 50 animals. The animals were maintained at 25 °C, transferred to fresh medium every day, and the number of dead flies was counted. Lifespan experiments were repeated three times.

Espadas J., Pendin D., Bocanegra R., Escalada A., Misticoni G., Trevisan T., Velasco del Olmo A., Montagna A., Bova S., Ibarra B., Kuzmin P.I., Bashkirov P.V., Shnyrova A.V., Frolov V.A, & Daga A. (2019). Dynamic constriction and fission of endoplasmic reticulum membranes by reticulon. Nature Communications, 10, 5327.

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Genetic Tools for Drosophila Research

Cited 2 times

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All UAS-sona RNAi lines and sona mutants generated in our lab and characterization of sona-Gal4 line (P{GawB}CG9850) are described^{28 (link)}. P{lacW}Diap1^j5C8/TM3 was used as Diap1-lacZ line^{57 (link),58 (link)}. UAS-Diap1, UAS-lacZ, rpr-lacZ, wg-lacZ, UAS-GFP, UAS-p35, hsFLP; FRT42D ubiGFP, FRT42D ubi-GFP, ptc-Gal4, en-Gal4, tubulin-Gal4 and apterous-Gal4 were obtained from Bloomington stock center. UAS-rpr RNAi was obtained from VDRC (101234). ci-Gal4 is described in^{59 (link)}. Fly cultures were carried out at 25 °C unless otherwise indicated.

Tsogtbaatar O., Won J.H., Kim G.W., Han J.H., Bae Y.K, & Cho K.O. (2019). An ADAMTS Sol narae is required for cell survival in Drosophila. Scientific Reports, 9, 1270.

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Drosophila Genetic Manipulation Toolkit

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UAS-catalase, UAS-SOD1, UAS-SOD2, UAS-p38b⁺, UAS-DMKK3⁺, UAS-coracle⁺, Tubulin-GAL4, Armadillo-GAL4 (Arm-GAL4), and Tubulin-Gal80^ts were from the Bloomington Stock Center. UAS-catalase^RNAi, UAS-SOD1^RNAi, UAS-SOD2^RNAi, UAS-DMKK3^RNAi, and UAS-coracle^RNAi were from the Vienna Drosophila RNAi Center. D-p38a¹³ and D-p38b^156A were as previously described (Chen et al., 2010 (link)). UAS-p38b^DN (Adachi-Yamada et al., 1999 (link)) was a kind gift from T. Adachi-Yamada at Kobe University, Japan. Dot-GAL4 (insertion 11C (c2)) (Kimbrell et al., 2002 (link)) was previously generated in D.A. Kimbrell’s laboratory at the University of California, Davis. GMH5 was as previously described by Wessells et al. (2004) (link). Hand-GAL4 was a kind gift from A. Paululat (University of Osnabruek, Germany).

Lim H.Y., Wang W., Chen J., Ocorr K, & Bodmer R. (2014). ROS Regulate Cardiac Function via a Distinct Paracrine Mechanism. Cell reports, 7(1), 35-44.

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Drosophila Genetic Manipulation Protocols

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Flies were maintained on standard cornmeal-agar medium at 29°C unless otherwise indicated. dTet-GFP flies were kindly provided by Ruth Steward. Tubulin-Gal4 (#5138), Slit-Gal4 (#9580), and Sim-Gal4 (#9150) fly stocks were obtained from Bloomington Drosophila Stock Center. dTet-RNAi (#102273 and #36187) and mCherry-RNAi (#35785) flies were obtained from Vienna Drosophila RNAi Center.

Ismail J.N., Badini S., Frey F., Abou-Kheir W, & Shirinian M. (2019). Drosophila Tet Is Expressed in Midline Glia and Is Required for Proper Axonal Development. Frontiers in Cellular Neuroscience, 13, 252.

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