Bp clonase kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The BP Clonase kit is a molecular biology tool used for performing site-specific recombination reactions. It facilitates the transfer of DNA fragments between entry and destination vectors. The kit contains the necessary enzymes and buffers to enable efficient and reliable cloning.

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Lab products found in correlation

Gateway cloning, by Thermo Fisher Scientific (1 mentions) Pdonr221, by Thermo Fisher Scientific (1 mentions) Glufosinate, by Bayer (1 mentions) Tcs sp2, by Leica (1 mentions)

Spelling variants of this product

BP clonase (133 citations) BP Clonase II kit (5 citations) BP and LR clonase kits (2 citations) Gateway BP Clonase II Enzyme Kit (2 citations) Gateway BP Clonase™ II Enzyme Mix kit (2 citations) BP Clonase Reaction Kit (2 citations) BP&LR Clonase II kit (2 citations)

3 protocols using bp clonase kit

Generation and Selection of Transgenic Plants

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All oligonucleotide primers used in this study were synthesized by Integrated DNA Technologies. Transgenic plants were generated using the floral dip method (Clough and Bent, 1998 (link)) with the Agrobacterium tumefaciens strain EHA105. N-terminal and C-terminal GFP and RFP fusions with AP4E under control of the UBQ10 promoter were created in pUBN-Dest and pUBC-Dest vectors (Grefen et al., 2010 (link)) using Gateway™ cloning (Invitrogen). The AP4E CDS in pLIC6, obtained from ABRC clone DKLAT1G31730, was moved into pDONR221 (Invitrogen) using the BP Clonase kit. Stop codon insertion and frame correction for pUBN constructs was accomplished with primer pairs GR6&7 and GR8&9, respectively, before using the LR Clonase kit (Invitrogen) to move respective AP4E constructs into pUBN and pUBC destination vectors.
T1 seedlings from pUBN/pUBC transformed plants were sown on soil and selected by spray application of a 120 μg/ml Glufosinate (Liberty Herbicide, Bayer Crop Sciences) water solution containing 0.05% (v/v) Silwet-77 surfactant 7, 9, 12, and 14 days after germination.

Dahhan D.A., Reynolds G.D., Cárdenas J.J., Eeckhout D., Johnson A., Yperman K., Kaufmann W.A., Vang N., Yan X., Hwang I., Heese A., De Jaeger G., Friml J., Van Damme D., Pan J, & Bednarek S.Y. (2022). Proteomic characterization of isolated Arabidopsis clathrin-coated vesicles reveals evolutionarily conserved and plant-specific components. The Plant Cell, 34(6), 2150-2173.

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CYP83A1 and PAD3 Overexpression

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The full-length CYP83A1 coding sequence (CDS) without the stop codon was amplified by PCR from Col-0 cDNA and inserted into the Gateway vector pDONR207 using a BP Clonase kit (Invitrogen) to create a pDONR207-CYP83A1 CDS entry clone. Then an LR Clonase kit (Invitrogen) was used to introduce the inserts into the pEarleyGate 101/103 destination vector containing a 35S promoter and C-terminal HA/GFP fusion (Earley et al., 2006 (link)). The same methods were used to construct PAD3 overexpression constructs.

Liu S., Bartnikas L.M., Volko S.M., Ausubel F.M, & Tang D. (2016). Mutation of the Glucosinolate Biosynthesis Enzyme Cytochrome P450 83A1 Monooxygenase Increases Camalexin Accumulation and Powdery Mildew Resistance. Frontiers in Plant Science, 7, 227.

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In vivo Subcellular Localization of NnNAC Proteins

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ProtComp v9.0 (http://linux1.softberry.com/berry.phtml) was used to obtain the subcellular localization predictions of NnNAC proteins. Coding regions of NnNAC genes were cloned into entry vectors (pDONR221 Zeo) using high fidelity primers (Supplementary Table S1) following the BP-clonase kit instruction manual (Lot#2335893, Invitrogen by Thermo Fisher Scientific, United States). Transformed plasmids were then cloned into PMDC43 vectors with LR-clonase according to the manufacturer’s instructions. These plasmids were subsequently inserted into the Agrobacterium tumefaciens cv. GV3101 using electric shock method. Young leaves of tobacco plants (4–6 weeks-old) were selected for transformation assay according to the methods of Kokkirala et al. (2010) (link). A confocal laser scanning microscope (Leica TCS SP2; Leica microsystems, Wetzlar, Germany) was used to photograph the agroinfiltrated leaves 48 h after infiltration.

Song H., Liu Y., Dong G., Zhang M., Wang Y., Xin J., Su Y., Sun H, & Yang M. (2022). Genome-Wide Characterization and Comprehensive Analysis of NAC Transcription Factor Family in Nelumbo nucifera. Frontiers in Genetics, 13, 901838.

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