2.4 Subcellular localization
The full length coding sequence of OsLHY was fused to green
fluorescent protein (GFP) in the pM999 vector under the control of the
CaMV 35S promoter. Meanwhile, the RPL1 coding sequencing was used to
fuse with cyan fluorescent protein (CFP), as RPL1 can be used as
a nuclear marker (Zhang, Yuan, & Zhang, 2012). Then the two plasmids
were co-transformed into the rice etiolated stems protoplasts isolated
from ten-day-old Zhong Hua11, which was grown in the growth chamber at
28 °C under dark conditions. The method for rice protoplasts
transformation was based on the previously study (Bart, Chern, Park,
Bartley, & Ronald, 2006). Fluorescence signals were detected by an
Olympus FluoView FV1000 confocal microscope.