scholarly journals Fusion of Green Fluorescent Protein with the Zeocin™-Resistance Marker Allows Visual Screening and Drug Selection of Transfected Eukaryotic Cells

BioTechniques ◽  
1998 ◽  
Vol 24 (3) ◽  
pp. 478-482 ◽  
Author(s):  
Robert P. Bennett ◽  
Cindy A. Cox ◽  
James P. Hoeffler
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Eukaryotic Cells ◽  
Drug Selection ◽  
Resistance Marker ◽  
Visual Screening ◽  
Green Fluorescent ◽  
Selection Of

Motif trap: a rapid method to clone motifs that can target proteins to defined subcellular localisations

1999 ◽  
Vol 112 (23) ◽  
pp. 4207-4211
Author(s):  
L.A. Bejarano ◽  
C. Gonzalez
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Eukaryotic Cells ◽  
Subcellular Localisation ◽  
Dna Fragments ◽  
Rt Pcr ◽  
Rapid Procedure ◽  
Target Proteins ◽  
Protein Motifs ◽  
Green Fluorescent

We have developed a rapid procedure termed Motif Trap (MT) to identify protein motifs that are able to target proteins to a distinct subcellular localisation in eukaryotic cells. By expressing random DNA fragments fused to green fluorescent protein (GFP), individual cells with the GFP localisation of interest are readily isolated allowing for the expressed DNA fragments to be cloned by RT-PCR. These can then be used to identify the corresponding full-length cDNAs. Using MT, we have identified patterns of GFP localisation which correspond to every major organelle and compartment. We have shown that MT is useful to identify new sequences that determine subcellular localisation as well as known targeting motifs.

scholarly journals Genetic transformation of Coniochaeta sp. 2T2.1, key fungal member of a lignocellulose-degrading microbial consortium

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Nancy N Nichols ◽  
Ronald E Hector ◽  
Sarah E Frazer
Keyword(s):  
Green Fluorescent Protein ◽  
Genetic Transformation ◽  
Pure Culture ◽  
Ecological Niche ◽  
Fluorescent Protein ◽  
Microbial Consortium ◽  
Transformed Cells ◽  
Antibiotic Selection ◽  
Green Fluorescent ◽  
Selection Of

Abstract Coniochaeta sp. strain 2T2.1 is a key member of a microbial consortium that degrades lignocellulosic biomass. Due to its ecological niche and ability to also grow in pure culture on wheat straw, protocols for transformation and antibiotic selection of the strain were established. Hygromycin was found to be a reliable selectable transformation marker, and the mammalian codon-optimized green fluorescent protein was expressed and used to visualize fluorescence in transformed cells of strain 2T2.1.

scholarly journals Selection of co-transformed Dendrobium Sonia 17 using hygromycin and green fluorescent protein

2011 ◽  
Vol 55 (3) ◽  
pp. 572-576 ◽  
Author(s):  
C. S. Tee ◽  
M. Maziah ◽  
C. S. Tan ◽  
M. P. Abdullah
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Green Fluorescent ◽  
Selection Of

Green fluorescent protein as a visual selection marker for coffee transformation

Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Manoj Mishra ◽  
Santosini Devi ◽  
Alex McCormac ◽  
Nigel Scott ◽  
DongFang Chen ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Selectable Marker ◽  
Material Selection ◽  
Visual Selection ◽  
Agrobacterium Mediated Transformation ◽  
Low Level ◽  
Coffee Plants ◽  
Green Fluorescent ◽  
Selection Of

AbstractThe green fluorescent protein (GFP) was used as a visual selectable marker to produce transgenic coffee (Coffea canephora) plants following Agrobacterium-mediated transformation. The binary vector pBECKS 2000.7 containing synthetic gene for GFP (sgfp) S65T and the hygromycin phosphotransferase gene hph both controlled by 35S cauliflower mosaic virus CaMV35S promoters was used for transformation. Embryogenic cultures were initiated from hypocotyls and cotyledon leaves of in vitro grown seedlings and used as target material. Selection of transformed tissue was carried out using GFP visual selection as the sole screen or in combination with a low level of antibiotics (hygromycin 10 mg/L), and the efficiency was compared with antibiotics selection alone (hygromycin 30 mg/L). GFP selection reduced the time for transformed somatic embryos formation from 18 weeks on a hygromycin (30 mg/L) antibiotics containing medium to 8 weeks. Moreover, visual selection of GFP combined with low level of antibiotics selection improved the transformation efficiency and increased the number of transformed coffee plants compared to selection in the presence of antibiotics. Molecular analysis confirmed the presence of the sgfp-S65T coding region in the regenerated plants. Visual screening of transformed cells using GFP by Agrobacterium-mediated transformation techniques was found to be efficient and therefore has the potential for development of selectable marker-free transgenic coffee plants.

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