Plant gene repair and insertion vector based on combination of CRISPR/Cas9 and BeYDV and construction method
The invention discloses a plant gene repair or insertion vector based on combination of CRISPR/Cas9 and BeYDV.The construction method comprises the following steps that 1, a target sequence or an exogenous gene is inserted between a left homologous arm and a right homologous arm of a target gene, a...
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Zusammenfassung: | The invention discloses a plant gene repair or insertion vector based on combination of CRISPR/Cas9 and BeYDV.The construction method comprises the following steps that 1, a target sequence or an exogenous gene is inserted between a left homologous arm and a right homologous arm of a target gene, a homologous recombination element (LIR-gene-SIR) is obtained, and homologous recombination can be conducted on one to two target genes; (2) inserting sgRNA of one or more target genes into the MCS; and (3) transferring the fragment obtained in the step (2) into a plant binary expression vector containing a Cas9 expression cassette. Homologous sequence repair is carried out at a target sequence under the action of a homologous recombination repair mechanism by using a homologous DNA sequence generated by mass replication of BeYDV. By utilizing the method, homologous recombination can be performed on two genes at the same time, and gene editing can be performed on other target genes, so that the purpose of performing |
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