Spurious localizations of diX-indigo microcrystals generated by the histochemical GUS assay

1994 ◽  
Vol 3 (3) ◽  
pp. 176-181 ◽  
Author(s):  
Jean-Claude Caissard ◽  
Anne Guivarc'h ◽  
Jacques Rembur ◽  
Abdelkrim Azmi ◽  
Dominique Chriqui
Keyword(s):  
Gus Assay ◽  
Histochemical Gus Assay

scholarly journals Analysis of MeEf1A6 Gene Promoter Activity with In-vitro and In-vivo using Transient and Stable Expression Techniques in Tobacco Plant (Nicotiana tabacum)

2021 ◽  
Vol 3 (1) ◽  
pp. 1-8
Author(s):  
Galih Gibral Andalusia ◽  
Sony Suhandono ◽  
Ima Mulyama Zainuddin
Keyword(s):  
Gene Expression ◽  
Manihot Esculenta ◽  
Tobacco Plant ◽  
Stable Transformation ◽  
Blue Color ◽  
Gus Assay ◽  
Tobacco Plants ◽  
Histochemical Gus Assay

The promoter is a part of the gene that functions in carrying out the gene expression, and its work activity becomes a matter of concern to ensure that expression works effectively. MeEF1A6 (Manihot esculenta Elongation Factor 1 Alfa - 6) is a promoter derived from cassava plants (Manihot esculenta). In previous studies, the MeEF1A6 promoter was successfully isolated, introduced, and characterized into the pBI121 plasmid, replacing the CaMV35S promoter. This study aims to analyze the activity of MeEF1A6 promoters in-vivo and in-vitro by using transient and transgenic techniques in tobacco plants. The pBI121 plasmid containing the MeEF1A6 promoter was introduced into Agrobacterium tumefaciens strain AGL1 and LBA4404. The promoter's work was then analyzed by the result of introducing it into the tobacco plant using the transient and stable transformation. The whole part of explants was used for transient study and tested in a minimum of two biological replicates. Sixty sheets of explant leaves that have been cut were used for stable transformation. The promoter work analysis was carried out with the GUS gene expression that integrated with the promoter with histochemical GUS assay. The transient produced a blue color in the roots, stems, and leaves on the whole repetition. The transverse incision in the stem shows the blue color on the epidermis and procambium tissue. Stable transformation using AGL1 as vector produced 43 shoots from 40 calli. A total of 43 shoots were selected with antibiotics and produced 27 plantlets that were successfully grown. Some plantlets are then reacted with x-gluc as histochemical GUS assay substrat and produced a blue color in the explants, indicating that the MeEF1A6 promoter has been successfully introduced. The results indicate that the MeEF1A6 promoter could work on plant tissue in roots, stems, leaves, and tissues that connect meristems such as procambium in tobacco plants. This reinforces the suspicion that the MeEF1A6 promoter performs work constitutionally as a constitutive promoter.   

scholarly journals Agrobacterium-mediated genetic transformation of two varieties of Brassica: optimization of protocol

1970 ◽  
Vol 34 (2) ◽  
pp. 287-301 ◽  
Author(s):  
MMA Khan ◽  
ABMAHK Robin ◽  
MAN Nazim-Ud-Dowla ◽  
SK Talukder ◽  
L Hassan
Keyword(s):  
Brassica Napus ◽  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Brassica Rapa ◽  
Binary Vector ◽  
Gus Assay ◽  
Key Words Transformation ◽  
Gus Reporter ◽  
Best Response ◽  
Transformation Experiment

 Two rapeseed varieties, namely Tori-7 and BARI Sarisha-8, respectively, from Brassica rapa and Brassica napus were selected to observe the transformation ability. Petioles were inoculated in Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector pBl2l with GUS (reporter) and nptII (kanamycin resistant) gene. The transformation experiment was performed by optimizing two important factors: preculture time and co-cultivation time and also selected out the best variety. Infection was most effective when explants were pre-cultured for 72 hours (80% GUS positive). and co-cultivated for 72 hours (72% GUS positive). The variety Tori-7 showed the best response to GUS assay (65% GUS positive). Callus induction was the highest in Tori-7, which were 6% with 72 hours of preculture period and 9% in 48 hours of co-cultivation. Number of putative transformed plantlets were highest in Tori-7 (7 plants) followed by BARI Sarisha-8 (3 plants). Key words: Transformation; Brassica; GUS; Agrobacterium. DOI: 10.3329/bjar.v34i2.5802Bangladesh J. Agril. Res. 34(2): 287-301, June 2009

Comparing the Effect of Two Promoters on Cassava Somatic Embryo at Transient GUS Assay Level

2015 ◽  
Vol 7 (2) ◽  
pp. 79-84
Author(s):  
Olufemi Oyelakin ◽  
Jelili Opabode ◽  
Emmanuel Idehen
Keyword(s):  
Somatic Embryo ◽  
Gus Assay

scholarly journals Genetic Transformation of Kentucky Bluegrass with the rolC Gene

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 616d-616 ◽  
Author(s):  
Shanqiang Ke ◽  
Chiwon W. Lee ◽  
Zong-Ming Cheng
Keyword(s):  
Southern Hybridization ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Naphthaleneacetic Acid ◽  
Gus Assay ◽  
Similar Frequency ◽  
Rolc Gene ◽  
Putative Transformants ◽  
Regenerated Plants ◽  
Npt Ii

Coleoptile tissues excised from young seedlings of `Touchdown' Kentucky bluegrass (Poa pratensis L.) were bombarded with the disarmed Agrobacterium tumefaciens strain EHA 101 carrying rolC (from A. rhizogenes), NPT II and GUS genes. These tissues were then cultured on Murashige and Skoog (MS) medium containing 0.2 mg·L–1 picloram, 0.01 mg·L–1 naphthaleneacetic acid (NAA), 150 mg·L–1 kanamycin, and 50 m acetosyringone. Calli formed on this medium within 2 weeks. The regenerated plants from these calli were analyzed for the presence of the GUS and rolC genes by histochemical GUS assay, PCR, and Southern hybridization. Only 3.7% of the regenerants were transformed when determined by the GUS assay. A similar frequency of transformation in the regenerated plants was obtained after bombarding the coleoptile tissues with the DNA isolated from the pGA-GUSGF-rolC plasmid. Most of the putative transformants were either albinos or variegated plants that are composed of both albino and green tissues.

scholarly journals Identification and characterization of a novel multi-stress responsive gene in Arabidopsis

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0244030
Author(s):  
Faiza Tawab ◽  
Iqbal Munir ◽  
Zeeshan Nasim ◽  
Mohammad Sayyar Khan ◽  
Saleha Tawab ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Survival Rates ◽  
Crop Productivity ◽  
Cell Wall Protein ◽  
Transcriptome Data ◽  
Gus Assay ◽  
Transgenic Lines ◽  
Responsive Gene ◽  
Identification And Characterization

Abiotic stresses especially salinity, drought and high temperature result in considerable reduction of crop productivity. In this study, we identified AT4G18280 annotated as a glycine-rich cell wall protein-like (hereafter refer to as GRPL1) protein as a potential multistress-responsive gene. Analysis of public transcriptome data and GUS assay of pGRPL1::GUS showed a strong induction of GRPL1 under drought, salinity and heat stresses. Transgenic plants overexpressing GRPL1-3HA showed significantly higher germination, root elongation and survival rate under salt stress. Moreover, the 35S::GRPL1-3HA transgenic lines also showed higher survival rates under drought and heat stresses. GRPL1 showed similar expression patterns with Abscisic acid (ABA)-pathway genes under different growth and stress conditions, suggesting a possibility that GRPL1 might act in the ABA pathway that is further supported by the inability of ABA-deficient mutant (aba2-1) to induce GRPL1 under drought stress. Taken together, our data presents GRPL1 as a potential multi-stress responsive gene working downstream of ABA.

Large-scale characterization of promoters from grapevine (Vitis spp.) using quantitative anthocyanin and GUS assay systems

Plant Science ◽  
2012 ◽  
Vol 196 ◽  
pp. 132-142 ◽  
Author(s):  
Zhijian T. Li ◽  
Kyung-Hee Kim ◽  
Jonathon R. Jasinski ◽  
Matthew R. Creech ◽  
Dennis J. Gray
Keyword(s):  
Large Scale ◽  
Gus Assay ◽  
Scale Characterization ◽  
Vitis Spp

β-Glucuronidase (GUS) Assay in Animal Tissue

Gus Protocols ◽  
1992 ◽  
pp. 189-203 ◽  
Author(s):  
John W. Kyle ◽  
Nancy Galvin ◽  
Carole Vogler ◽  
Jeffrey H. Grubb
Keyword(s):  
Animal Tissue ◽  
Gus Assay

scholarly journals Functional GUS assay of GRAS transcription factor from Medicago truncatula

2019 ◽  
Vol 33 (1) ◽  
pp. 1187-1194
Author(s):  
Miglena Revalska ◽  
Mariana Radkova ◽  
Lyuben Zagorchev ◽  
Anelia Iantcheva
Keyword(s):  
Transcription Factor ◽  
Medicago Truncatula ◽  
Gus Assay ◽  
Gras Transcription Factor

Application of the GUS marker gene technique to high-throughput screening of rhizobial competition

1999 ◽  
Vol 45 (8) ◽  
pp. 678-685 ◽  
Author(s):  
Kate J Wilson ◽  
Adriana Parra ◽  
Lina Botero
Keyword(s):  
Phaseolus Vulgaris ◽  
Sample Size ◽  
Microbial Ecology ◽  
High Throughput ◽  
High Throughput Screening ◽  
Marker Gene ◽  
Nodule Occupancy ◽  
Gus Gene ◽  
Gus Assay ◽  
Rhizobium Strains

The GUS marker gene system has been developed for the study of bacterial ecology, particularly rhizobial competition. For high-throughput field screening of rhizobial competition, the technique must be robust and reliable under diverse conditions, with diverse cultivars and strains. Here we demonstrate its applicability to the evaluation of competition on five different Phaseolus vulgaris cultivars with 10 different Rhizobium strains. We describe refinements of the GUS assay, which make it more affordable and applicable to field-based studies, and use the assay to examine the effect of sample size on the accuracy of nodule occupancy measurements.Key words: GUS gene, Rhizobium, rhizobial competition, microbial ecology.

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