Uas p35

Manufactured by BD

Sourced in United States

The UAS-p35 is a laboratory equipment designed for sample preparation and processing. It is a versatile device that can be used for a variety of applications in research and clinical settings.

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6 protocols using uas p35

Comprehensive Drosophila Genetic Toolkit

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The following stocks and crosses were used:
Fig. 1: sco/SM5; vg^QEGal4 (BDSC #8229). UAS-Transtimer/SM5; TubGal80ts(BDSC #7017)/TM6B. The UAS-Transtimer was provided by Li He (Norbert Perrimon's Lab, Harvard Medical School, MA, USA).
Fig. 2. vg^BEGal4 (BDSC #8222)/SM5; UAS-GPFn (BDSC #4776)/TM6B. UAS-vgRNAi (2nd chr.,) from Vienna Drosophila Resource Center #16896)/SM5; UAS-Dcr2/TM6B. UAS-p35 (BDSC #6298); UAS-vgRNAi/SM5, UAS-Dcr2 (BDSC #90938)/TM6B. UAS-hid (BDSC #65408)/SM5; MKRS, hs-Flp/TM6B.
Fig. 3 and Fig. S3, apGal4 (BDSC #56807), UAS-GPFn (BDSC #4775)/SM5; vgQElacz (3nd chr.,)/TM6B. UAS-vgRNAi/SM5, UAS-Dcr2 (BDSC #90938)//TM6B.
Fig. 4, hs-Flp; apGal4 (BDSC #56807), UAS-GPFn (BDSC #4775)/SM5; Tub-FRT-STOP-FRT-Gal80 (BDSC #5145)/TM6B. UAS-p35 (BDSC #6298); UAS-vgRNAi/SM5, UAS-Dcr2 (BDSC #90938)//TM6B.
Unless indicated otherwise, all flies and crosses were reared on standard culture medium at 25°C.

Flores-Flores M., Muñoz-Nava L.M., Rodríguez-Muñoz R., Zartman J, & Nahmad M. (2023). Vestigial-dependent induction contributes to robust patterning but is not essential for wing-fate recruitment in Drosophila. Biology Open, 12(5), bio059908.

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Fly Stocks Used in Genetic Studies

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Some stocks used in this study were kindly from Dr. Qing Zhang’s lab, including UAS-rdx-RNAi [33 (link)], UAS-HA-rdx [33 (link)], rdx-lacZ [33 (link)], UAS-Ub-K48R [32 (link)], UAS-Ub-K63R [32 (link)], UAS-Ub-K0 [32 (link)] and UAS-HA-rdx-Δ3box [32 (link)]. UAS-ci and rdx^Δ6 were gifted from Junzheng Zhang’s lab. UAS-ci-RNAi (NIG #2125R-1), UAS-P35 (BDSC #5072), UAS-smo (BDSC #44620), GMR-hid (BDSC #5771), Ay-gal4 (BDSC #4411), UAS-GFP (BDSC #1522), GMR-gal4 (BDSC #8605), MS1096 (BDSC #8860), UAS-Diap1 (BDSC #6657), Ap-gal4 (BDSC #3041) were obtained from NIG or BDSC. The detailed information of these fly stocks has been described in Flybase database. The tub-Myc-diap1 construct was made by cloning a full-length diap1 cDNA downstream of the α-tubulin promoter [53 (link)]. Then the tub-Myc-diap1 construct was injected into w¹¹¹⁸Drosophila embryos according to the method described previously [21 (link)].

Liu B., Ding Y., Sun B., Liu Q., Zhou Z, & Zhan M. (2021). The Hh pathway promotes cell apoptosis through Ci-Rdx-Diap1 axis. Cell Death Discovery, 7, 263.

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Drosophila Genetics Protocol

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All Drosophila strains were grown and maintained at 25 °C. Following fly stocks were used for the experiments: Ciao1 RNAi (Vienna Drosophila Resource Center, v32020 and v105939), Xpd RNAi (Vienna Drosophila Resource Center v106998), UAS-CycE (BDSC 4781), UAS-Diap1, UAS-p35, ey-Gal4, GMR-Gal4, en-Gal4, ptc-Gal4, nub-Gal4, and FRT42D M(2)53¹ (BDSC 5698) were obtained from the Bloomington stock center. For overexpression of Crb^intra, UAS-Crb^intra was crossed with GMR-Gal4 (Bloomington). Xpd^p flies were a kind donation from Dr. Beat Suter. To construct the transgenic lines, UAS-Ciao1 and UAS-Xpd, full length Ciao1 and Xpd cDNA (from the Drosophila Genome Research Center) were cloned into pUAST vector.

Jung J., Yeom E, & Choi K.W. (2020). Ciao1 interacts with Crumbs and Xpd to regulate organ growth in Drosophila. Cell Death & Disease, 11(5), 365.

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Drosophila Genetic Manipulation Protocol

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Flies were reared on a standard corn meal, yeast, and sucrose agar medium at 25°C under a 12-hour/12-hour light/dark cycle. Canton S (BDSC 64349), painless [EP(2)2451] (BDSC 27895), ppk-Gal4 (BDSC 32078), UAS-CD8-GFP (BDSC 5130), UAS-Dcr2, UAS-tetanus toxin (active, BDSC 28838 and inactive BDSC 28839), UAS-p35 (BDSC 5072), and UAS-Lamin-GFP (BDSC 7376) flies were obtained from the Bloomington Drosophila Stock Center (BDSC) library. w1118 (VDRC 60000), UAS-TrpA1-RNAi (VDRC 37249), UAS-RDL-RNAi (VDRC 41101), UAS-GRD-RNAi (VDRC 5329), UAS-D-GABA-B-R1-RNAi (VDRC 101440), UAS-D-GABA-B-R2-RNAi (VDRC 110268 and VDRC 1785), UAS-D-GABA-B-R3-RNAi (VDRC 50176), UAS-LCCH3-RNAi (VDRC 37408), UAS-Vglut-RNAi (BDSC 27538), UAS-ChAT-RNAi (BDSC 25856), UAS-nAChRα1-RNAi (VDRC 48159), and UAS-twist-RNAi (VDRC 37091 and VDRC 37092) flies were obtained from the Vienna Drosophila Resource Center (VDRC) RNAi library (66 (link)). UAS-TrpA1 and TrpA1^ins flies were from P. Garrity, and Gad1-Gal4 flies were from H. Bellen.

Khuong T.M., Wang Q.P., Manion J., Oyston L.J., Lau M.T., Towler H., Lin Y.Q, & Neely G.G. (2019). Nerve injury drives a heightened state of vigilance and neuropathic sensitization in Drosophila. Science Advances, 5(7), eaaw4099.

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Drosophila Genetic Tools and Strains

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All fly stocks were reared at 25°C on standard flour/agar Drosophila media. The Gal4/UAS system [58 (link)] was used to drive the expression of transgenes at 29°C. The following strains were provided by the Bloomington Drosophila Stock Center (BDSC) or the Vienna Drosophila RNAi Center (VDRC): Df(3R)BSC503 (BDSC 25007); Df(3R)ED6332 (BDSC 24141); hdc⁴³(BDSC 64063); hdc⁵⁰(BDSC 64064); hdc^BG23007(BDSC 12410); UAS-hdc (BDSC 64056); hdc^RI(VDRC 104322); hdc^R2(VDRC 45069); hdc^R3(BDSC 30489); UAS-p35 (BDSC 5073); UAS-Rheb (BDSC 9689); UAS-Xbp1-EGFP (BDSC 60731); Xbp1^RI(BDSC 36755); PEK^RI(VDRC 110278); UAS-myristoylated-Tomato (BDSC 32222); UAS-CD8-GFP (BDSC 5137); UAS-src-GFP (BDSC 5429); UAS-myrRFP (BDSC 7138); hsp70-GFP (BDSC 51354); phmGal4 (BDSC 80577); fkhGal4 (BDSC 78060); apGal4 (BDSC 3041), UAS-GFPnls (BDSC 4776); TM3-cherry (BDSC 35524); puc-LacZ (BDSC 11173); The following strains are described in Flybase: nub-Gal4 [59 (link)]; Ci-Gal4 [60 (link)]; sal^E/PV [61 (link)]; amn^c651 [10 (link)]. Gal4 drivers were recombined to UAS fluorescent markers described here. UAS-SOD1::UAS-Cat recombined construct was kindly provided by F. Serras. phmGal4::YPetAtet [16 (link)] was kindly provided by X. Franch-Marro. w¹¹⁸ strain was used as control.

Giannios P, & Casanova J. (2021). Systemic and local effect of the Drosophila headcase gene and its role in stress protection of Adult Progenitor Cells. PLoS Genetics, 17(2), e1009362.

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Genetic Manipulation of Drosophila for Research

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Drosophila stocks were maintained on a standard cornmeal medium. The following transgenic stocks were utilized in this study: P{w+; UAS-GFP::dPRL^WT}, P{w+; UAS-GFP::dPRL^C173S}, P{w+; GMR-RGH-miRNA} [34 (link)], P{w+; GMR-reaper-miRNA} [34 (link)], P{w+; GMR-grim-miRNA} [34 (link)], P{w+; GMR-hid-miRNA} [34 (link)], P{w+; UAS-dPRL-IR⁴⁵⁵¹⁸} (Vienna Drosophila Resource Center (VDRC, Vienna, Austria, stock No. 45518), GMR-Gal4 (Bloomington Drosophila Stock Center (BDSC, Bloomington, IN, USA, stock No. 1104), UAS-dicer2 (BDSC, stock No. 24646), P{w+; UAS-p35} (BDSC, stock No. 5072). To ensure the GAL4 expression and efficiency for RNA interference, flies were raised at 27 °C for pupal dissection. The wild-type coding sequence of dPRL were subcloned into the pUAST vector containing a GFP coding sequence upstream of the cloning site for transgenes. The QuikChange Lightning Site-Directed Mutagenesis kit (Agilent Technologies, Santa Clara, CA, USA) was used to generate the Cys173 to Ser substitution in the dPRL coding sequence. Transgenic flies were generated according to the standard protocol via microinjection.

Chen S.F., Hsien H.L., Wang T.F, & Lin M.D. (2023). Drosophila Phosphatase of Regenerating Liver Is Critical for Photoreceptor Cell Polarity and Survival during Retinal Development. International Journal of Molecular Sciences, 24(14), 11501.

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