Boea clarkeane drought-resistant and salt-tolerance related gene and its coding protein and use
A technology of Capsulatum, coding protein, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as lack of stress-resistant resource genes, and achieve the effect of improving drought tolerance and high practical application value
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Embodiment 1
[0030] Example 1, Acquisition of Drought Resistance and Salt Tolerance Related Gene BhLea1 of Capsulatum
[0031] Total RNA was extracted from the leaves of Capsulatum spp. leaves subjected to drought stress for 8 hours, and a cDNA library was constructed using the ZAP-cDNA(R) library construction kit (Stratagene, La Jolla, CA). 4800 genes were randomly selected from them, and automatically sampled on Hybond-N with BioGrid robot (BioRobotics Ltd, Cambridge, UK). +cDNA microarrays (cDNA chips) were fabricated on nylon membranes (Amersham Biosciences, Freiburg, Germany). The polyA-RNA prepared by combining the chip with the polyA-RNA of the leaves of Capsula oleracea that grew normally without drought treatment and the leaves of Capsula capsulae leaves that were drought-stressed for 8 hours 33 P-labeled probes were hybridized to analyze their dynamic changes in gene expression under normal conditions and after drought induction. The genes after drought induction were sequenced...
Embodiment 2
[0032] Example 2, BhLea1 expression pattern analysis experiment
[0033] Extract the total RNA of the leaves of Capsulatum spp. without stress treatment, drought stress for 2, 8, 24, and 72 hours, and 0.2M NaCl stress and flooding stress for 9 hours, and measure the concentration. Take equal amounts of each sample for 1% agarose gel electrophoresis, after the bromophenol blue-stained sample migrates to the 2 / 3 of the gel away from the sample hole, transfer the RNA to the nylon membrane, bake the membrane at 80°C for 120 minutes, and then transfer the RNA to the nylon membrane. The nylon membrane of the above RNA was put into a hybridization tube, and after pre-hybridization at 65°C for 2.5 h, the purified and denatured fluorescein-labeled BhLea1 probe (from 1 to 588 at the 5' end of SEQID No. 1 in the sequence table) was added. Base position nucleotide sequence), hybridize at 65°C for 24 hours, pour out the hybridization solution, wash the membrane with 2×SSC, 0.1% SDS, 0.1×SS...
Embodiment 3
[0034] Example 3 Cytological localization of BhLea1
[0035] Using PSORT computer program analysis software (Nakai and Kanehisa, Expert system for predicting protein localization sites in gram-negative bacteria. Proteins. 11 (2): 95-110, 1991) to carry out cytological localization of the protein BhLea1 encoded by BhLea1, the results It indicated that BhLea1 was localized in the cytoplasm of the leaf cells of Capsula oleifera.
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