Agroinfiltration protocol
WebFor transient expression, agroinfiltration is a simple and effective method to deliver transgenes into plant cells. After an Agrobacterium infection, recombinant proteins can … Plasmids were mobilized into A. tumefaciens strains AGL1, C58C1 and LBA4404 via electroporation [36]. Recombinant agrobacteria were prepared for infiltration using a modified protocol of Sainsbury and Lomonossoff [32]. In short, a single colony of recombinant bacteria was inoculated into liquid LB media (10 g/L … See more pEAQ-HT was a generous gift from Sainsbury and Lomonossoff, John Innes Centre, UK [32]. p35S-GSN is a pBIN-Plus vector backbone containing the uidA gene (with a small … See more The 35S-CMV.2b (1-94)-nos gene cassette, encoding the C terminal CMV 2b truncation, was amplified from p35S-CMV.2b (1-94) using primers 35S_FseI-F and nosT_FseI-R … See more N. benthamiana leaf samples were collected between 0 and 8 days post agroinfiltration and snap frozen in liquid nitrogen. Total soluble protein (TSP) was extracted by … See more Chemicals including lipoic acid (0–100 µM; Merck), ascorbic acid (0–100 mM; Merck), PVP (0–1 g/L; Merck) and Pluronic F-68 (0–0.2%; Thermo Fisher Scientific) were filter sterilized and added to the MMA/bacteria mix … See more
Agroinfiltration protocol
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WebApr 5, 2024 · In this study, we developed an improved cowpea protoplast isolation protocol, a transient protoplast assay, and an agroinfiltration assay to be used for initial testing … WebJun 23, 2024 · This protocol combines a vacuum-based Agroinfiltration approach and the high translational efficiency of a deconstructed virus-based binary vector (pEAQ-HT). …
WebApr 10, 2024 · Sclerotinia sclerotiorum is a broad host range necrotrophic fungal pathogen, which causes disease on many economically important crop species.S. sclerotiorum has been shown to secrete small effector proteins to kill host cells and acquire nutrients. We set out to discover novel necrosis-inducing effectors and characterize their activity using … WebMar 7, 2024 · Total protein was extracted 48 h post-agroinfiltration and incubated with anti-Flag magnetic beads. VvBKI1-GFP can be detected by Western blotting ( Figure 2 B). Furthermore, we performed GST pull-down assay with glutathione S-transferase (GST)-tagged VvAPX1 and His-tagged VvBKI1 synthesized from Escherichia coli (DE3).
WebProtocol DOI: 10.1385/1-59745-003-0:225. Affiliations: Department of Plant Pathology, University of Arkansas, Fayetteville, AR ... Two days after agroinfiltration, Arabidopsis plants can be treated with chemical inducers such as dexamethasone to induce transgene expression. The expression of the β-glucoronidase (GUS) reporter gene can be ... Web23 hours ago · 3.2 Subcellular localization of MirMAN protein. The marker for plasma membrane (PM), PAD62-mcherry, was co-expressed with pCAMBIA1303-35S-MirMAN-GFP by Agrobacterium-mediated transient expression in Nicotiana benthamiana leaves. After 16 h of agroinfiltration, we observed fluorescence in the green channel with a confocal …
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WebApr 3, 2024 · Agroinfiltration is a valuable research method for investigating virulence and avirulence effector proteins from pathogens and pests, where heterologous effector proteins are transiently expressed in plant leaves and hypersensitive necrosis responses and other effector functions can be assessed. substitute for ezo lower denture cushionsWebIn the present study, we report a tobacco syringe agroinfiltration and liquid chromatography tandem mass spectrometry (TSAL)-based method for performing a phytohormone transporter activity assay using endogenous hormones present in tobacco (Nicotiana benthamiana) leaves. ... The step-by-step protocol of the TSAL method is … paint chip repair tool carWebProtocol 1. Plant Growing and Testing Conditions Grow and maintain plants in controlled greenhouses or climate chambers within the temperature range of 18-22 °C and under natural light regime or with a 16 hr/8 hr day/night regime. Remove axillary branches in order to make the plants more manageable. substitute for elizabethan collarWebAgroinfiltration is conducted by infiltration of agrobacterial suspension (0.4 OD) into intercellular spaces of fingernail-sized leaves that are still attached to the intact plant. A … paint chip repair near me come to your homeWebMar 19, 2024 · The GFP protein was expressed transiently in lettuce, Nicotiana benthamiana, tomatoes, eggplants, hot peppers, melons, and orchids with agroinfiltration. Compared to a single terminator, a double ... paint chip sample sizeWebDay 1: Agrobacteriumtransformation with binary vectors. Day 3: Start Agrobacteriumculture (including Agrobacterium containing p19 strain).p19 is a suppressor of gene silencing from the tomato bushy stunt virus and must be coinfiltrated … substitute for egg whites in royal icingWebSep 21, 2024 · A genetically engineered chimeric virus crTMV-CP-PLRV composed of the crucifer-infecting tobacco mosaic virus (crTMV) RNA and the potato leafroll virus (PLRV) coat protein (CP) was obtained by agroinfiltration of Nicotiana benthamiana with the binary vector pCambia-crTMV-CPPLRV. The significant levels of the chimeric virus enabled … substitute for farmers cheese in pierogies