Plant salt resistant associated protein AtST1, its coded genes and application
A technology encoding genes and proteins, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problem of unclear gene function, and achieve the effect of improving hypertonicity and ABA resistance.
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Embodiment 1
[0042] Example 1. Acquisition of proteins and genes encoding them in response to high-salt stress in plants
[0043] Total RNA was extracted from Arabidopsis thaliana (Columbia type, also known as Col-0 ecotype) seedlings (purchased from the Arabidopsis Biological Resource Center (ABRC)) seedlings for 10 days, and the RNA was digested with DNase from Takara Company. The digested DNA was reverse-transcribed to obtain cDNA. Using the cDNA as a template, a pair of primers respectively provided with XbaI and KpnI restriction sites were used for PCR amplification to obtain the full-length cDNA of salt-tolerance-related genes. The primer sequences are as follows:
[0044] Primer1: 5'-tctaga ATGGAATCGAGTAGCGTTGATGAG-3' (XbaI);
[0045] Primer2: 5'-ggtaccTTACGAGGCGTGAAAGATGCGTTGC-3' (KpnI).
[0046] The high-fidelity Pyrobest DNA polymerase from Takara Company was used for amplification. The amplification system was 50 μL, containing 5.0 μL of 10×Pyrobest PCR buffer, 1.0 μL of 10 mM ...
Embodiment 2
[0053] Example 2. Functional verification of proteins and genes encoding them in response to high-salt stress in plants
[0054] 1. Obtaining AtST1 overexpression plants (Super: AtST1-1 and Super: AtST1-2)
[0055] 1. Intermediate vector pCAMBIA1300: Construction of Super
[0056] Utilize primers aagcttGTGGGCCTGTGGTCTCAAGAT and tctagaCTAGAGTCGATTTGGT from plasmid pMSP-1 (GenBank number is EU181145) to amplify the Super promoter of the Super promoter whose nucleotide sequence is shown in the 113-1112 position from the 5' end of sequence 3 in the sequence listing Sequence; the amplified Super promoter was double-digested with HindIII and XbaI, and the Super promoter fragment was recovered and inserted between the HindIII and XbaI restriction sites of plasmid pCAMBIA1300 (GenBank No. FJ362601) to obtain pCAMBIA1300: Super plasmid.
[0057] 2. Super: Construction of AtST1
[0058] The recombinant vector pMD18-AtST1 containing the AtST1 gene obtained in Example 1 and the interme...
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