Agroinfiltration protocol
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 WebJun 5, 2009 · We demonstrated that the agroinfiltration-based transient gene expression can be used to detect potato proteins in sub-cellular compartments in living cells. We …
Agroinfiltration protocol
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WebNational Center for Biotechnology Information WebJul 28, 2024 · Infiltration has found a special application in plant biotechnology as a method of transient transformation with the use of Agrobacterium suspension (agroinfiltration) …
WebFeb 14, 2024 · An Improved Syringe Agroinfiltration Protocol to Enhance Transformation Efficiency by Combinative Use of 5-Azacytidine, Ascorbate Acid and Tween-20. Plants … WebAgroinfiltration 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 …
WebAug 25, 2024 · Background: Agroinfiltration is a simple and effective method of delivering transgenes into plant cells for the rapid production of recombinant proteins and has … 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 ...
WebOur Forest trees have crucial economic press ecological value. As a model tree, poplar has played a significant role in elucidating the molecular mechanisms underlying tree biology. However, an defect in mutant my and time-consuming barn genetic transformation processing severely limit progress into the functional characterization of asian genes. A …
WebIn 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 … hcfa 1500 place of serviceWebAgroinfiltration 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 needleless plastic syringe is used to infiltrate bacterial suspension into the underside of leaves ( see Note 5 ). 2. hcfa 1500 or ubWebThis protocol combines a vacuum-based Agroinfiltration approach and the high translational efficiency of a deconstructed virus-based binary vector (pEAQ-HT). The described methodology is robust, easy to perform, and results in high amount of transient expression in C. roseus. This protocol is expected to serve as valuable tool to enhance … gold coast diving toursWeb23 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 … gold coast djWebProtocol 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. gold coast dog foodWebApr 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 … gold coast doggy daycareWebJan 1, 2009 · Agroinfiltration has been used successfully in many different applications, including the analysis of small RNAs. We describe here a protocol for analysis of miRNA processing using agroinfiltration of N. benthamiana leaves. Key words Agroinfiltration Transient assay miRNA processing Small RNA blot Download protocol PDF hcfa 1500 place of service codes