Application method of stress tolerance related gene ZmHDZIV14 in regulation and control of plant stress resistance
A technology for plant stress resistance and related proteins, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, and other methods of inserting foreign genetic materials. and other problems, to achieve the effect of improving stress resistance
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Embodiment 1
[0040] Embodiment 1, the acquisition of ZmHDZIV14 protein and its coding gene
[0041] Seeds of wild-type Arabidopsis thaliana (Arabidopsisthaliana, Col) were preserved by Gansu Key Laboratory of Maize in Arid Habitat Crop Science.
[0042] Bacterial strain: Escherichia coli (Eschrichiacoli) DH5a, Agrobacterium tumefaciens (Agrobacterium tumefaciens) used for genetic transformation of Arabidopsis thaliana is LBA4404, both strains are preserved by Gansu Provincial Key Laboratory of Crop Science in Arid Habitat.
[0043] Plasmid: The T / A cloning vector pUCm-T was purchased from Sangon Bioengineering (Shanghai) Co., Ltd., the plant expression vector pCAMBIA3300-35S-PROIIMCS-bar was transformed by the Gansu Key Laboratory of Crop Science in Arid Habitats, and Kanna resistance was used for T / A cloning.
[0044] Embodiment 1; The acquisition of SEQIDNO:2 protein and its coding gene
[0045] Bacterial strains: Escherichia coli (Eschrichiacoli) DH5a, Agrobacterium tumefaciens (Agrob...
Embodiment 2
[0074] Example 2; Functional verification of ZmHDZIV14 and its coding gene
[0075] One, the construction of ZmHDZIV14 gene expression vector
[0076] (1) The target fragment is connected to the pUCm-T vector
[0077] Configure the following solution according to the instructions of pUCm-Tvector (Sangon Biology): 10×LigationBuffer 1.0 μl, 50% PEG 1.0 μl, pUCm-Tvector 1.0 μl, purified PCR product 4.0 μl, T4DNALigase 1.0 μl, ddH 2 O2 μl. Ligation at 16°C for 6 hours.
[0078] (2) Preparation of Escherichia coli competent cells (prepared in 1.5ml centrifuge tube)
[0079] Transfer the bacterial solution into a pre-cooled 1.5ml centrifuge tube, centrifuge at 3500rpm at 4°C for 10min, and remove the supernatant. Add 300 μl of ice-cold 0.1 mol / L CaCl2 solution to the centrifuge tube to resuspend the bacteria, and ice-bath for 30 minutes. Centrifuge at 3500 rpm for 10 min at 4°C. Remove the supernatant, suspend the bacteria in 60 μl of ice-cold 0.1 mol / L CaCl2 solution, add 10 ...
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