Gt115

Manufactured by InvivoGen

The GT115 is a laboratory equipment product manufactured by InvivoGen. Its core function is to facilitate the measurement of gene expression levels in cells or tissues.

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Lab products found in correlation

Triple layer flasks, by Thermo Fisher Scientific (1 mentions) Celline cl 350 bioreactor flasks, by Integra LifeSciences (1 mentions) Hybridoma sfm medium, by Thermo Fisher Scientific (1 mentions) Hitrap qff columns, by GE Healthcare (1 mentions) Heparin hp column, by GE Healthcare (1 mentions)

Close spelling variants of this product

GT115 bacteria E. coli GT115 cells Escherichia coli GT115 Chemically competent E. coli GT115

3 protocols using gt115

Vibrio fischeri Strains and Culture Conditions

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The bacterial strains used in this study are listed in Table 1 and were derived from ES114, a wild-type
V. fischeri strain isolated from E.
scolopes (Boettcher & Ruby,
1990). V. fischeri derivatives were generated using
previously described conjugation (Visick &
Skoufos, 2001) mutagenesis (Le Roux
et al., 2007, Shibata et al., 2012 (link)), and transposon
(Tn7) chromosomal-insertion (McCann et al., 2003 (link)) methods.
V. fischeri cells were grown in Luria-Bertani Salt (LBS)
media (Graf et al.,
1994), Seawater Tryptone (SWT) media (Boettcher & Ruby, 1990 (link)), or HEPES Minimal Media (HMM) (Ruby & Nealson, 1977 (link)). E.
coli strains used for molecular genetics in this study include:
ER2508 (NEB), TAM1 λ pir (Active Motif), π3813
(Le Roux et al.,
2007), CC118 λ pir (Herrero et al., 1990 (link)), and GT115
(Invivogen). E. coli strains were grown in LB (Davis et al., 1980 ). Solid media
contained 1.5% agar. For V. fischeri, antibiotics were
added to the following concentrations when necessary: erythromycin (Erm) at 5
μg ml⁻¹, tetracycline (Tet) at 5 μg
ml⁻¹ in LBS or 30 μg ml⁻¹ in
SWT and HMM, or chloramphenicol (Cm) at 2.5 μg ml⁻¹.
The following antibiotics were added to E. coli media where
appropriate: Cm at 25 μg ml⁻¹, Tet at 15 μg
ml⁻¹, kanamycin (Kan) at 50 μg
ml⁻¹, or ampicillin (Ap) at 100 μg
ml⁻¹.

Norsworthy A.N, & Visick K.L. (2015). Signaling between two interacting sensor kinases promotes biofilms and colonization by a bacterial symbiont. Molecular microbiology, 96(2), 233-248.

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Vibrio fischeri Bacterial Strains and Plasmids

Cited 1 time

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V. fischeri strains and plasmids used in this study are listed in Table 1, and additional details of their construction are located in Supporting Information. All V. fischeri strains were derived from wild-type strain ES114 (Ruby et al. 2005 (link); Mandel et al. 2008 (link)). Escherichia coli strains used in this work include EC100Dpir+ (Epicentre Biotechnologies, Madison, WI), TAM1 (Active Motif, Carlsbad, CA), β3914 (Le Roux et al. 2007 (link)), π3813 (Le Roux et al. 2007 (link)), CC118 λpir (Herrero et al. 1990 (link)), and GT115 (InvivoGen, San Diego, CA). Oligonucleotides used in this study are listed in Table S1 and were purchased from Integrated DNA Technologies, Inc. (Coralville, IA) (IDT).

Miyashiro T., Oehlert D., Ray V.A., Visick K.L, & Ruby E.G. (2014). The putative oligosaccharide translocase SypK connects biofilm formation with quorum signaling in Vibrio fischeri. MicrobiologyOpen, 3(6), 836-848.

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Purification and Characterization of Recombinant Human PCSK9

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The sequence of human PCSK9 used here is identical to sequence of GenBank acc. no. CAC38896.1. The coding region was ligated into the pCpGfree-vitroNmcs expression vector and transformed into the E. coli strain GT115 encoding the pir gene (Invivogen). CHO-K1 cells stably transfected with PCSK9 were grown as a suspension in Hybridoma-SFM medium (GIBCO) and then expanded to Celline CL 350 Bioreactor flasks (Integra) or in triple layer flasks (Thermo Fischer Scientific). The medium from PCSK9-expressing CHO cells was 1:1 in 25 mM Tris pH 7.4 and applied to two serial-connected 5 mL HiTrap QFF columns (GEhealthcare). The columns were washed with 25 mM Tris pH 7.4 and protein subsequently eluted with 25 mM Tris pH 7.4 and 1 M NaCl. The eluted sample was dialyzed against 25 mM Tris pH 7.4 and 5% (v/v) glycerol at 4°C. The dialyzed sample was applied to a 5 mL Heparin HP column (GEhealthcare) and protein was eluted in 1 mL fractions by a linear gradient over 40 mL from 0 to 1 M NaCl in 25 mM Tris pH 7.4. Fractions containing PCSK9 were concentrated and further purified by SEC on a Superdex200 increase 10/300 or 16/600 in either PBS or 25 mM Tris pH 7.4 and 150 mM NaCl. Samples were analyzed by SDS-PAGE and PCSK9 was concentrated to 5–8 mg/mL based on absorbance at 280 nm and flash-frozen in liquid nitrogen before storage at-80°C.

Pärn A., Olsen D., Tuvikene J., Kaas M., Borisova E., Bilgin M., Elhauge M., Vilstrup J., Madsen P., Ambrozkiewicz M.C., Goz R.U., Timmusk T., Tarabykin V., Gustafsen C, & Glerup S. (2023). PCSK9 deficiency alters brain lipid composition without affecting brain development and function. Frontiers in Molecular Neuroscience, 15, 1084633.

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