Plant drought resistance related protein OsERF62, coding gene and application thereof
A drought-tolerant, plant technology, applied in the direction of plant genetic improvement, application, plant peptides, etc., can solve the problems of slow progress, great chance, unstable varieties, etc.
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Embodiment 1
[0046] Example 1. Acquisition of Plant Drought Tolerance-Related Protein OsERF62 and Its Encoding Gene
[0047] The upland rice IRAT109 seedlings cultivated under normal conditions were taken, the total RNA was extracted by Trizol method, and after purification, the cDNA was obtained by reverse transcription with M-MLV reverse transcriptase. Using the cDNA as a template, primers F1 and R1 were designed for PCR amplification, the amplified product was subjected to agarose gel electrophoresis, and a DNA fragment of 1580 bp was recovered and purified for sequencing. The result is shown in sequence 2 of the sequence table.
[0048] The sequences of the above primers are as follows:
[0049] F1: 5′-CGG GGTACC AAAGGCATTCGCAACACACA-3′ (the underlined base is the restriction endonuclease KpnI restriction endonuclease recognition sequence);
[0050] R1: 5′-CC TTAATTAA CCAAAATACATTACGACTGGAC-3' (the underlined base is the recognition sequence for restriction endonuclease PacI).
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Embodiment 2
[0052] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of the endogenous OsERF62 gene
[0053] 1. Coercion treatment
[0054] The rice variety Nipponbare and the upland rice variety IRAT109 seedlings with a seedling age of 4 weeks were subjected to the following treatments respectively:
[0055] 1. ABA treatment: Soak the roots of the seedlings in 100 μM ABA solution, culture in light for 1 hour, 2 hours, 4 hours, 6 hours, 9 hours, 12 hours, and 36 hours, then take the leaves, and freeze them with liquid nitrogen at -80°C Save for later.
[0056] 2. Dehydration treatment: put the seedlings in the air for 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, and 8 hours, then take the leaves, freeze them with liquid nitrogen, and store them at -80°C for later use.
[0057] 3. H 2 o 2 Treatment: Soak the roots of the seedlings in 1mM H 2 o 2 After being placed in the solution for 1 hour, 2 hours, 4 hours, 6 hours, 9 hours, 12 hou...
Embodiment 3
[0065] Embodiment 3, the construction of recombinant expression vector
[0066] After the DNA fragment of 1580bp obtained in Example 1 was double-digested with KpnI and PacI, the digested product was connected to the backbone fragment of the vector pMDC32 double-digested by KpnI and PacI, and confirmed by sequencing and digestion, the recombinant vector 35S was obtained: :OsERF62, the recombinant vector 35S::OsERF62 is inserted between the KpnI and PacI sites of the vector pMDC32 (that is, the downstream of the double tobacco mosaic virus promoter 35S) from the 10th to the 1570th nucleotide of the sequence table sequence 2 DNA fragments shown in sequence.
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