Application of OsWRKY53 in forward regulation of BR signal
A positive regulation and signaling technology, applied in the application, introduction of foreign genetic material using vectors, angiosperms/flowering plants, etc., can solve problems such as unclear mechanism of action
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[0047] Example 1. Obtainment of OsGSK2-RNAi oswrky53 double mutant
[0048] 1. Input the CDS sequence of the OsWRKY53 gene into the CRISPR Primer Designer software, design 2 pairs of target site primers (F1 and R1; F2 and R2), and construct the knockout vector CRISPR / Cas9-OsWRKY53.
[0049] Forward primer F1: 5'-GGCATTCCAGTCGTACCCTCTGAGC-3'
[0050]Reverse primer R1: 5'-AAACGCTCAGAGGTACGACTGGAA-3'
[0051] Forward primer F2: 5'-GCCGAGCTGGAGGACGGGTACAAC-3'
[0052] Reverse primer R2: 5'-AAACGTTGTACCCGTCCTCCAGCT-3'
[0053] 2. Using OsGSK2-RNAi transgenic rice as experimental material, the OsGSK2-RNAi oswrky53 double mutant was obtained by Agrobacterium-mediated genetic transformation.
[0054] 3. Design a pair of sequencing primers (F3 and R3) covering the target sequence, amplify the target sequence, and sequence the PCR product until a homozygous OsGSK2-RNAi oswrky53 double mutant is obtained.
[0055] Forward primer F3: 5'-CGGGGTGCCCAAGTTCAAGTC-3'
[0056] Reverse prime...
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