Bj5183 competent cells

Manufactured by Agilent Technologies

Sourced in United States

The BJ5183 competent cells are a specialized bacterial strain used in molecular biology and genetic engineering applications. These cells are designed to facilitate the manipulation and cloning of DNA fragments. The core function of the BJ5183 competent cells is to enable efficient transformation and propagation of plasmid DNA, which is a crucial step in various DNA-based experiments and genetic engineering workflows.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Padeasy 1 vector, by Agilent Technologies (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Padeasy, by Agilent Technologies (1 mentions) Adeasy adenoviral vector system, by Agilent Technologies (1 mentions)

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BJ5183 cells

3 protocols using bj5183 competent cells

Adenoviral Expression of IRF6

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Adenovirus production followed the AdEasy protocol56 (link). Briefly, human IRF6 cDNA from pCMV-SPORT6-IRF6 plasmid was cloned into pAdTrack-CMV vector (Agilent Technologies) at Xho I and Xba I restriction enzyme sites. The AdIRF6 adenovirus expression plasmid was generated from recombination between pAdEasy-1 vector and pAdTrack-CMV-IRF6 in BJ5183 competent cells (Agilent Technologies). AdIRF6 plasmid (40 μg) was linearized with Pac I restriction enzyme and subsequently transfected into AD293 cells using Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer’s protocol for adenovirus packaging. Adenoviruses were harvested at 14 to 20 days after transfection.

Ke C.Y., Xiao W.L., Chen C.M., Lo L.J, & Wong F.H. (2015). IRF6 is the mediator of TGFβ3 during regulation of the epithelial mesenchymal transition and palatal fusion. Scientific Reports, 5, 12791.

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Adenoviral Transduction of HER2 Mutants

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Plasmid #16259 (human HER2, V654E) and plasmid #16258 (human HER2, K753M) were purchased from Addgene and sequenced. Both plasmids were originally in the vector pcDNA3 with 5’ and 3’ HindIII cloning sites. They were cloned into the HindIII site of pAdTrack-CMV (He et al., 1998 (link)). These constructs were recombined with pAdEasy in BJ5183 competent cells (Agilent Technologies). Subsequently, Adenovirus5 (Ad5) was produced and titrated using standard methods. Western blot experiments were used to verify the activity of the constructs, as described in the Results section. The GFP virus was purchased from Vector Biolabs. All viral protocols were reviewed by the URMC Biosafety committee to ensure the safety of staff and the environment.

Zhang J., Wang Q., Abdul-Aziz D., Mattiacio J., Edge A.S, & White P.M. (2018). ERBB2 signaling drives supporting cell proliferation in vitro and apparent supernumerary hair cell formation in vivo in the neonatal mouse cochlea. The European journal of neuroscience, 48(10), 3299-3316.

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Recombinant Adenovirus Production

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The AdEasy Adenoviral vector system (Agilent Technologies, Santa Clara, CA, USA Catalog #240031 and #240032) was used to generate recombinant adenoviruses. Full length cDNA of human WT or S75Y RagC was subcloned into the pshuttle vector, pshuttle–IRES–hrGFP under the control of CMV promoter. The constructed vector was co-transformed into BJ5183 competent cells (Agilent Technologies, Santa Clara, CA, USA) together with pAdEasy-1, the viral DNA plasmid. Recombinant Adenovirus plasmid DNA was generated via homologous recombination and then transfected AD293 cells with lipofectamine 2000 (Thermo Fisher, Waltham, MA, USA) to produce recombinant adenoviruses, Ad:RagC^WT and Ad:RagC^S75Y. The Ad:GFP was used as a control.

Kim M., Lu L., Dvornikov A.V., Ma X., Ding Y., Zhu P., Olson T.M., Lin X, & Xu X. (2021). TFEB Overexpression, Not mTOR Inhibition, Ameliorates RagCS75Y Cardiomyopathy. International Journal of Molecular Sciences, 22(11), 5494.

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