The full-length ORF of the AaMYB17 gene without the terminator codon was inserted into the pHB-YFP expression vector under the CaMV35S promoter to form a pHB-AaMYB17–YFP fusion protein. Then the plasmid and p19 protein were introduced into the A. tumefaciens strain GV3101 for N. benthamiana leaf transient expression (Sparkes et al., 2006 (link)). The fluorescent signals were observed 60–72 h after infiltration using a TCS SP5-II confocal laser microscopy (Leica Microsystems, Wetzlar, Germany). Three biological repeats were performed to verify these results.
Tcs sp5 2 confocal laser microscopy
Manufactured by Leica
Sourced in Germany
The TCS SP5-II is a confocal laser microscopy system manufactured by Leica. It is designed to provide high-resolution imaging of biological samples through the use of laser excitation and confocal detection. The system features advanced optics, sensitive detectors, and intuitive software for capturing and analyzing images.
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2 protocols using tcs sp5 2 confocal laser microscopy
1
Subcellular Localization of AaMYB17
2
Transient Expression of AaNAC1-YFP Fusion
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The ORF of AaABF3 gene without the terminator codon TAG was inserted to the PHB-YFP expression vector under the CaMV35S promoter to form a PHB-AaNAC1–YFP fusion protein. The plasmid and P19 protein were then transformed into A. tumefaciens strain GV3101 for transient expression in the epidermal cells of N. benthamiana. Agrobacterium cells were cultured in MS liquid medium to an OD600nm of 0.6 and injected into leaves of 6-week-old N. benthamiana after incubation at room temperature for 3 h. The YFP signals were imaged 60 h later using a TCS SP5-II confocal laser microscopy (Leica Microsystems, Germany).
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