2024 Phee401e vector

Phee401e vector

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WebDec 19, 2024 · To construct pHEE401E (Wang et al., 2015) vectors targeting four different sites of PAR1 and one site of JAZ1 or GAI, the dsDNA was fused to the BsaI-digested … WebNov 22, 2024 · The pHEE401E vector has a type IIS restriction-enzyme-recognition site for ligation [6]. The other is a pBAtC-tRNA vector that uses a polycistronic tRNA-gRNA …

4: Egg cell-specific promoter-controlled CRISPR/Cas9 binary …

WebSep 24, 2024 · The structure of CSE-653 vector designed to sequentially cut S653 codon twice. pHEE401E backbone was used to ubiquitously express two sgRNAs by U6-26 and U6-29 promoters, and Cas9 was driven by an ... WebJul 24, 2024 · By Gibson Assembly, this element was ligated into the pHEE401E binary vector (Wang et al., 2015) digested by restriction endonuclease BsaI. We added the BsaI restriction site to the GL2-2Tar-F/R primers, so the final vector still retained the BsaI cutting site, allowing for the insertion of sgRNAs for target loci of interest. diwali fashion for men

Addgene: pHSE401

WebJan 7, 2024 · pHEE401E article : bacterial resistance : Kanamycin cloning : growth notes : Please see the cloning protocol attachments for detailed descriptions on how to insert sgRNA sequences into this plasmid. growth strain : Egg cell-specific promoter-controlled expression of 3 FLAG-NLS-zCas9-NLS. WebpHSE401 (Plasmid #62201) Print Enlarge View all sequences Purpose CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA … WebI am trying to synthesize CRISPR constructs, using Golden Gate to put my sgRNAs in the pHEE401E vector. Whenever I do colony PCR with my original primers, it appears that the reaction worked as I... diwali fashion show theme

Study of vacuole glycerate transporter NPF8.4 reveals a new role …

WebThe construct is split into one module vector set (pCBC-DT1T2) containing two single guide RNAs (sgRNAs) and a binary vector based on pCAMBIA (pHEE401E) containing additional … WebpHEE401E . Cat# PEV-141 ... Description. A plant dual expression plasmid. Vector Type. Plant Dual Expression Vector, CRISPR . Expression Host. Plant Cell . Form. Powder; Solution Size. 2 μg; 10 μL . High Copy/Low Copy. High Copy ... craftsman weedwacker air filterWebGenetic transformation (GT) has emerged as a powerful tool for exploration of plant-RKN interactions and genetic improvement of RKN resistance. However, it is usually difficult to achieve a highly efficient and stable GT protocol for most crops due to the complexity of this process. Results diwali festival facts for kids

"WebNov 22, 2024 · The pHEE401E vector has a type IIS restriction-enzyme-recognition site for ligation [6]. The other is a pBAtC-tRNA vector that uses a polycistronic tRNA-gRNA expression system and has two AarI type IIS restriction enzyme sites. Originally, these two vectors could carry two sgRNAs. " - Phee401e vector

Phee401e vector

WebModified from the pHEE401E T-DNA vector described by Wang &al. (2015) Genome Biol 16: 144, doi:10.1186/s13059-015-0715-0: the Hygromycin selection maker of the T-DNA was exchanged with a cassette enabling selection on the basis of CFP-fluorescence in seeds; the egg-cell specific promoter of Cas9 was exchanged with a ubiquitin promoter. WebJan 20, 2024 · Agrobacterium cells harboring the pHEE401E plasmid were grown in Luria–Bertani medium for 2 d and then resuspended with 10 mL liquid MS medium. Disks (1 × 1 cm) were cut from N. benthamiana leaves using a scalpel and soaked in the Agrobacterium solution for 5 min. Leaf disks were then removed and placed onto the MS0 …

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WebNov 22, 2024 · The vectors used were pHEE401E_UBQ_Bar and pBAtC_tRNA, which employ a one-promoter/one-sgRNA and a polycistronic-tRNA-gRNA strategy, respectively. Golden Gate cloning was performed with type IIS... WebAug 16, 2024 · Building on the pHEE401E plasmid, a Cambia T-DNA adapted for 96 genome editing with CRISPR/Cas9 nucleases (Wang & al., 2015), we created a series of 97 vectors enabling selection and counter-selection of transgenics on the basis of seed 98 fluorescence. Toward this end, we replaced the hygromycin resistance marker of pHEE401E

WebPrinciple and design of JMJ epigenome editing constructs targeting APX2. Schematic representation of generated constructs for validation of sgRNA design by transient transactivation in N. benthamiana (A) and for generation of stable lines (B) in the pHEE401E vector backbone. Both types of constructs contain an sgRNA cassette, which is … WebDownload scientific diagram 4: Egg cell-specific promoter-controlled CRISPR/Cas9 binary vector pHEE401E: Two sgRNA expression cassettes pHEE401E vector was used to create …

WebJan 17, 2024 · pHEE401E binary vector. pHEE401E is based on pCambia backbone for Agrobacterium-mediated transformation. It contains the complementary parts of the … WebApr 18, 2015 · /dnas_title="pHEE401E" ORIGIN : 1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac: 61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga: 121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc: 181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc

WebNov 22, 2024 · The pHEE401E_UBQ_Bar vector is a version in which the egg-specific promoter for Cas9 expression in the pHEE401E vector was replaced with the UBQ10 …

WebJun 22, 2024 · a A schematic representation of the gene drive that is inserted into the second exon of the CRYPTOCHROME 1 ( CRY1) gene on Chromosome IV using CRISPR/Cas9-mediated homology-directed repair (HDR).... diwali festival holidays 2022WebApr 22, 2024 · After immunoprecipitation (IP) with anti-FLAG agarose, ubiquitinated BIK1 was detected by immunoblot (IB) using anti-HA antibodies (lanes 1 and 2) or treated with GST–USP2-cc (lane 3).... diwali festival activities for toddlersWebName. pHEE401E. Type. plasmid. Tair Accession. Vector:6531224345. Description. CRISPR/Cas9 vector with maize codon-optimized Cas9 between egg cell-specific enhancer/promoter combination (EC1.2en/EC1.1p) and rbcS-E9 terminator. Host Strain. diwali faral backgroundWebsgRNA sequence and cloned into pHEE401E binary vector, resulting in the production of pHEE401E-BUPS. With aim at knockouting RALF4 and RALF19 at the same time, we … diwali festival images downloadWebAug 13, 2024 · EAR (Ethylene-responsive element binding factor-associated Amphiphilic Repression) motif-containing transcription repressors have been shown to regulate plant growth and development, and plant responses to plant hormones and environmental stresses including biotic and abiotic stresses. However, the functions of most EAR-motif … craftsman weedwacker 4 cycle partsWebNational Center for Biotechnology Information craftsman weedwacker attachmentsWebSep 27, 2024 · The target-specified guide RNA (gRNA) sequences were cloned into a binary vector harbouring the cas9 gene under control of a ubiquitin promoter from parsley (Figure S1; Kawalleck et al ., 1993 ). The binary vectors targeting CsGTR1 and CsGTR2 harboured four gRNAs, while the vector for CsMYB28 and CsMYB29 contained three gRNAs. craftsman weedwacker 4 cycle manual