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Recombination gateway lr clonase enzyme mix kit

Manufactured by Thermo Fisher Scientific
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The Recombination Gateway LR clonase Enzyme mix kit is a laboratory tool designed for the recombination of DNA sequences. It facilitates the transfer of DNA fragments from an entry clone to an expression clone through an enzymatic process. The core function of this kit is to enable efficient and rapid cloning of DNA segments into various vector systems.

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Lab products found in correlation

Quickchange 2 kit, by Agilent Technologies (1 mentions) Pentr1a, by Thermo Fisher Scientific (1 mentions) Pentrtm1a, by Thermo Fisher Scientific (1 mentions)

2 protocols using recombination gateway lr clonase enzyme mix kit

1

Cloning and Mutagenesis of NLRP3

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Human NLRP3 were cloned in pENTR1A (Invitrogen) from Flag-NLRP3 encoding plasmids kindly shared by F. Martinon (University of Lausanne, Lausanne, Switzerland). Mutations were performed using QuickChange II kit (Agilent Technologies). cDNAs were transferred in pInducer21 using recombination Gateway LR clonase Enzyme mix kit (Thermo Fisher Scientific) (Meerbrey et al., 2011 (link)). pMD2.G and pCMVR8.74 were gifts from D. Trono (plasmid #12259 and #22036; Addgene).
Cosson C., Riou R., Patoli D., Niu T., Rey A., Groslambert M., De Rosny C., Chatre E., Allatif O., Henry T., Venet F., Milhavet F., Boursier G., Belot A., Jamilloux Y., Merlin E., Duquesne A., Grateau G., Savey L., Jacques Maria A.T., Pagnier A., Poutrel S., Lambotte O., Mallebranche C., Ardois S., Richer O., Lemelle I., Rieux-Laucat F., Bader-Meunier B., Amoura Z., Melki I., Cuisset L., Touitou I., Geyer M., Georgin-Lavialle S, & Py B.F. (2024). Functional diversity of NLRP3 gain-of-function mutants associated with CAPS autoinflammation. The Journal of Experimental Medicine, 221(5), e20231200.
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2

Cloning and Mutagenesis of NLRP3 Proteins

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Constructs coding for Flag-NLRP3 and Flag-NLRP3-LRR were a kind gift from F. Martinon (Unil, Lausanne, Switzerland), VSV-SGT1 from V. Petrilli (CRCL, Lyon, France), Myc-BRCC3 from W. Harper (Harvard Medical School, Boston), pcDNA3.1-HA-CSNK1A1 from P. Wang (Tongji University, Shanghai); pcDNA3-N-HA-NEK7 was purchased from Addgene. Human NLRP3 (hNLRP3), mouse NLRP3 (mNLRP3), and truncation deletions were cloned in pENTRTM1A (Invitrogen). Mutations were performed using QuickChangeTM II kit (Agilent Technologies). Primers are listed in Supplemental Table 3. cDNAs were transferred in pInducer21 using recombination Gateway LR clonase Enzyme mix kit (Thermofisher)38 (link).
Niu T., De Rosny C., Chautard S., Rey A., Patoli D., Groslambert M., Cosson C., Lagrange B., Zhang Z., Visvikis O., Hacot S., Hologne M., Walker O., Wong J., Wang P., Ricci R., Henry T., Boyer L., Petrilli V, & Py B.F. (2021). NLRP3 phosphorylation in its LRR domain critically regulates inflammasome assembly. Nature Communications, 12, 5862.
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