fluorescent protein egfp gene  (Addgene inc)

Journal: bioRxiv

Article Title: From Pigments to Precision: Exploring Genetic Transformation and Genome Editing in Wheat and Barley

doi: 10.1101/2024.12.03.626565

Figure Lengend Snippet: Protoplasts were transformed with linearized DNA by PEG-mediated transformation (top images); embryos were transformed with circular DNA by biolistic transformation (bottom images). Promoters activity was evaluated by monitoring GFP fluorescence intensity using a fluorescent microscope. Scale bar in upper and lower images = 100 and 500 µm, respectively.

Article Snippet: We linked each promoter to the wheat codon-optimized enhanced green fluorescent protein ( eGFP ) gene (Figure S2a) and cloned the constructs into the pJET1.2 vector (Thermo Fisher Scientific).

Techniques: Transformation Assay, Activity Assay, Fluorescence, Microscopy

Journal: Frontiers in Plant Science

Article Title: Fine-tuning CRISPR/Cas9 gene editing in common bean ( Phaseolus vulgaris L.) using a hairy root transformation system and in silico prediction models

doi: 10.3389/fpls.2023.1233418

Figure Lengend Snippet: (A) Position of the sgRNAs targeting the raffinose synthase ( PvRS1 and PvRS2 ) and stachyose synthase ( PvSS ) genes in P. vulgaris . (B) Schematic representation of the T-DNA of the pMR356-GFP and pMR394-GFP vectors. The A. thaliana codon-optimized Cas9 gene ( Atco-Cas9 ) was driven by a Parsley Ubiquitin promoter (PcUbi) in pMR356-GFP whereas Atco-Cas9 was driven by a 2X35S-Ω promoter in pMR394-GFP. In both vectors, the Atco-Cas9 gene was fused to a nuclear localization signal derived from simian virus 40 (SV40 NLS) and transcription was terminated by a A. thaliana heat shock protein 18.2 terminator (AtHSP18.2 ter). The transcription of the sgRNA was driven by a U6 promoter. The enhanced green fluorescent protein ( eGFP ) gene was placed under the regulation of a Cauliflower Mosaic Virus 35S promoter (CaMV 35S) and nopaline synthase terminator (NOS ter).

Article Snippet: For this study, both vectors were further improved by the incorporation of an enhanced green fluorescent protein gene ( eGFP ), under the control of a 35S promoter and terminated by a nopaline synthase terminator (NOS ter) with polyadenylation signal (transcription unit originating from the pGFPGUSPlus vector; RRID: Addgene_64401), using the NEBuilder HiFi DNA Assembly master mix (New England Biolabs, Ipswich, Massachusetts, United States of America).

Techniques: Ubiquitin Proteomics, Derivative Assay, Virus