Rice salt-tolerant gene OsRR22 mutant, encoded amino acid sequence thereof, plant and making method of mutant
A salt-tolerant gene and mutant technology, applied in the field of plant biology, can solve the problems of lack of gene editing and improvement reports, and achieve the effect of maintaining comprehensive agronomic traits and improving salt tolerance
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[0030] This example relates to the acquisition of rice OsRR22 gene knockout mutants, that is, the creation method.
[0031] 1. Guide RNA target sequence design and selection
[0032] According to the principle of controlling the genome sequence of the rice salt-tolerant gene OsRR22 and the design target site of CRISPR-Cas9 technology, the present invention designs the target site on the first exon of the OsRR22 gene, designs, selects and synthesizes one OsRR22 For guide RNA target sequences, see figure 1 and Table 1.
[0033] Table 1 Oligonucleotide sequences of gRNA targets
[0034] gRNA target sequence
Oligonucleotide sequence (5'-3')
RR22-F
ggcAGAGGGATCAATTCCCCGT
RR22-R
aaacACGGGGAATTGATCCCCTCT
[0035] 2. Construction of CRISPR / Cas9-gRNA vector
[0036] Referring to the method of Ma et al. (Ma et al., 2015, Molecular Plant, DOI: 10.1016 / j.molp.2015.04.007), take an equal amount of upstream and downstream primers (Table 1) of the ...
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