Detection of target sites of CRISPR/Cas9 system in melon by Agrobacteri-um rhizogenes system
The cultivation melon Long Qing No.8 was used as the receptor material to construct the CRISPR/Cas9 edit-ing vector of CmCURT1A gene.The targe site was detected through Agrobacterium rhizogenes,which provided the vector basis for subsequent genetic transformation experiment of melon.A double target knockout vector was constructed using the CmCURT1A gene(ID:MELO3C006053.2)as the target gene.Through a simple genetic transformation technique me-diated by Agrobacterium rhizome K599,adventitious roots were grown.PCR sequencing revealed that different base frag-ments of 65 bp and 72 bp were absent in the adventitious roots.The method was simple and efficient for the detection of CRISPR/Cas9 vector target site knockout in melon,realizing rapid identification of gene editing targets,and laying a foun-dation for studying the gene function and genetic improvement in melon.
万方数据
维普
