The overexpression vector MdHY5-pCAMBIA1300 was constructed by inserting the MdHY5 open reading frame into the transformed vector pCAMBIA1300.
To generate MdHY5 transgenic apple calli, the recombinant plasmid was introduced into Agrobacterium tumefaciens LBA4404 as described by An et al.36 (link) Regarding the transformation of apple calli, 15-day-old ‘Orin’ calli (WT) were co-cultured with Agrobacterium carrying MdHY5-pCAMBIA1300. The calli were co-cultured on MS medium containing 1.5 mg L−1 2, 4-dichlorophenoxyacetic acid and 0.5 mg L−1 6-butyric acid for 2 days at room temperature. The calli were then washed three times with sterile water and transferred to selective media supplemented with 300 mg L−1 carbenicillin and 35 mg L−1 hygromycin for transgene selection. The transgenic apple calli were co-cultivated in selective media that contained appropriate concentrations of antibiotics.
To generate MdHY5 transgenic apple calli, the recombinant plasmid was introduced into Agrobacterium tumefaciens LBA4404 as described by An et al.36 (link) Regarding the transformation of apple calli, 15-day-old ‘Orin’ calli (WT) were co-cultured with Agrobacterium carrying MdHY5-pCAMBIA1300. The calli were co-cultured on MS medium containing 1.5 mg L−1 2, 4-dichlorophenoxyacetic acid and 0.5 mg L−1 6-butyric acid for 2 days at room temperature. The calli were then washed three times with sterile water and transferred to selective media supplemented with 300 mg L−1 carbenicillin and 35 mg L−1 hygromycin for transgene selection. The transgenic apple calli were co-cultivated in selective media that contained appropriate concentrations of antibiotics.
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