Greengate cloning

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August undefined, 2024

WebOct 12, 2024 · The hip1 coding sequence devoid of BsaI restriction sites for GreenGate cloning was synthesized by Biocat (Germany). For recombinant protein expression, the hip1 sequence was amplified without SP and cloned into the pET28a (+) expression vector (Addgene, USA), eventually having a His-tag at the N-terminus. Sequences for cysteine …

Development and analysis of a highly flexible multi-gene

WebNow, a new toolkit - aptly named GreenGate – offers all of those same advantages to researchers working in plant model systems. “The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann of Heidelberg University. “What is different is the plasmids you use to bring this into your target organism. WebOct 15, 2024 · One cloning kit, termed GreenGate, was designed to facilitate rapid cloning of . constructs using commonly used elements of plant transformation vectors [16]. Assembly . dewitt city wide garage sale

Plasmids 101: Golden Gate Cloning - Addgene

WebCloning is performed by pipetting in a single tube all plasmid donors, the recipient vector, a type IIS restriction enzyme and ligase, and incubating the mix in a thermal cycler. … WebHome - PLOS WebDec 20, 2013 · GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules (plant promoter, N-terminal tag, coding sequence, C-terminal tag, plant terminator and plant resistance cassette), but at the same time allows assembling several expression cassettes in one … dewitt cleaners dewitt ny

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Figure 1. The Golden Gate principle. A) Type IIS restriction...

WebDec 19, 2013 · A) The GreenGate cloning system uses six different types of pUC19 based entry vectors into which the individual elements are inserted and a pGreen-IIS based destination vector. Magenta scissors represent BsaI recognition sites. In each GreenGate reaction, six modules are ligated between the left border (LB) and the right border (RB) … WebApr 27, 2024 · An important prerequisite for a molecular toolkit is a universal and cost-effective cloning method that can be easily adopted by different laboratories. To this end, we generated a set of plasmids based on the GreenGate cloning system (Lampropoulos et al., 2013; Figure 1; Supplemental Table S1). This Golden Gate cloning system allows … dewitt city police department miWebThis plasmid is designed for Golden Gate cloning using the BsaI enzyme. These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were … dewitt city hall iowa

"WebJan 20, 2024 · Here, we adapted the GreenGate cloning vectors to create two destination vectors showing strong marking of cell membranes in either the whole root or specifically in the lateral roots. This system can also be used in both embryos and whole seedlings. As proof of concept, we follow both gene expression and anatomy in Arabidopsis … " - Greengate cloning

Greengate cloning

A molecular toolkit for the green seaweed Ulva mutabilis

WebThis plasmid is designed for Golden Gate cloning using the BsaI enzyme. Plasmid Features (listed as bp in full plasmid sequence): BsaI site #1 = 2241-2251bp D-dummy = 2252-2297bp ... GreenGate - A Novel, Versatile, and Efficient Cloning System for Plant Transgenesis. Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner J ... WebThe characteristic of GreenGate cloning technology is sufficient to simultaneously construct a variety of effector expression vectors fused with different tags. On the basis of the …

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Webday 1: amplify lvl0 unit, like promoter, terminator etc., perform GG reaction, transform; day 2: check E.coli by PCR, seed correct clones; day 3: prepare the plasmid, which can be … WebGolden Gate cloning is one of the easiest cloning methods in terms of hands-on time, as digestion and ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a …

Web57 rows · GreenGate - A Novel, Versatile, and Efficient Cloning System for Plant Transgenesis. Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner J … WebJul 19, 2024 · The driver lines were generated employing the fast and flexible GreenGate cloning system (Lampropoulos et al., 2013) but are compatible with any vector/transgenic line in which the expression of an effector is under the control of derivatives of the pOp promoter element (Moore et al., 1998). An important feature of our driver lines is the ...

WebOct 29, 2024 · Cloning Greengate entry module construction. Primers with GreenGate overhangs were designed following the Greengate protocol. Control primers were designed at the same time for colony screens and sequencing. For a list of primers see Supplemental Table S1. Construction of destination vectors with red membrane included WebJul 12, 2024 · GreenGate - a novel, versatile, and efficient cloning system for plant transgenesis. PLoS One 8 , e83043 (2013). Article ADS Google Scholar

WebDec 20, 2013 · GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of …

WebNational Center for Biotechnology Information church revitalization conferenceWebGreenGate - A Novel, Versatile, and Efficient Cloning System for Plant Transgenesis. Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner J PLoS One. 2013 Dec 20;8(12):e83043. doi: 10.1371/journal.pone.0083043. PubMed Plasmids from Article ... dewitt cleaners incWebThe characteristic of GreenGate cloning technology is sufficient to simultaneously construct a variety of effector expression vectors fused with different tags. On the basis of the abundant six modules provided by GreenGate toolkit, it is also possible to use empty modules to carry out modifications for specific research. The availability of ... church revival flyerWebAug 9, 2015 · The development of DNA cloning technologies and the discovery of Agrobacterium sp. as a vehicle for delivery of recombinant DNA into the chromosomes of … church revivalWebGreenGate Cloning System to create plant expression vectors containing several cassettes and generate multi-construct transgenic plants - Jan Lohmann. MoClo Plant Parts II and Infrastructure Kit for hierarchical assembly of plant multigene constructs - … dewitt cleanersWebFor this study, plasmid constructs were generated via GreenGate cloning . The protein-coding regions of MsSND1, ... For ectopic expression of transcription factors, the respective GreenGate construct was transformed into Agrobacterium tumefaciens ASE (pSOUP +). Selected transgenic clones were incubated in liquid LB medium containing the ... church revival bulletin coversWeband versatile cloning system for the generation of plant transfor-mation vectors, which we named GreenGate. The GreenGate system allows rapid and efficient assembly of six … dewitt cleaners hours