Greengate cloning

Author: jmnj

August undefined, 2024

WebGreenGate cloning system for plant transgenesis. Developed by Jan Lohmann and colleagues, GreenGate is a cloning system for the rapid assembly of plant transformation constructs. As the name suggests, GreenGate is based on the Golden Gate cloning method, but has been modified specifically to improve plant transgenesis. 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 ...

Dual expression and anatomy lines allow simultaneous …

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 … 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 … devonshire pottery marks

Addgene: GreenGate - A Novel, Versatile, and Efficient …

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 . 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 … 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 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 ... churchill victoria postcode

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GreenGate cloning technology introduces enormous advantages …

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 … WebThis 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 … churchill vic weather forecastWebFor 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 ... devonshire practice portsmouth

"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 " - Greengate cloning

Greengate cloning

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 … 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. …

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WebJul 12, 2024 · GreenGate - a novel, versatile, and efficient cloning system for plant transgenesis. PLoS One 8 , e83043 (2013). Article ADS Google Scholar 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) …

WebHome - PLOS WebThe GreenGate cloning method was employed to generate the complemented lines 73. To this end, the coding sequence of DEIP1 was amplified by PCR with primers cl_AT2G27290_For and cl_AT2G27290_Rev. ...

Webdestination vector for GreenGate cloning method, contains 2x a set of modules from A-F, "Driver line", tissues-specific promoter, Dex-inducible mTurquoise2 expression: WebDec 20, 2013 · GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of …

WebThe plasmids used in this study were generated using either standard molecular cloning procedures (Sambrook et al., 1989) or by using the GreenGate Cloning system (Lampropoulos et al., 2013 ...

WebMay 14, 2024 · The Golden Gate cloning method is an easy and straight forward procedure. Here, Lampropoulos et al. describe a variety of it designed for plant transgenesis, calling it cleverly GreenGate. The whole procedure is possible due to type IIS restriction enzymes. These enzymes cut at a defined number of nucleotides after the … churchill victoria weather forecastWebGreenGate 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 - … devonshire pool tableWebA) 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 … devonshire preschool chatsworthWebJan 4, 2024 · Understanding the context-specific role of gene function is a key objective of modern biology. To this end, we generated a resource for inducible cell-type specific trans-activation based on the well-established combination of the chimeric GR-LhG4 transcription factor and the synthetic pOp promoter. Harnessing the flexibility of the GreenGate … churchill victoria populationWebOct 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 … churchill view my documentsWebGreenGate Cloning Kit Description: GreenGate is a simple and efficient cloning system for rapidly assembling plant transformation constructs. It is based on the Golden Gate … devonshire primary academy blackpoolWebGreenGate cloning technology is sufficient to simultaneously construct a variety of effector expression vectors. All these make GreenGate versatile and applicable cloning system … churchill victoria australia