2.5 Subcellular Localization
The full-length coding sequences of CsHSP24.6 and CspTAC5lacking its termination codon were separately cloned into the entry
vector pDONR207 using the Gateway BP Enzyme mix (Invitrogen) with
specific primers. The plasmids newly extracted after sequencing were then
introduced into the destination vector named pGWB5 [fused with green
fluorescent protein (GFP) at the C-terminal] using the LR Clonase
enzyme (Invitrogen). After sequencing, the plasmids
pGWB5-CsHSP24.6 and pGWB5-CspTAC5 were separately
transformed into the Agrobacterium tumefaciens strain GV3101 as
described above. Selection of a positive colony for cultivation and
infiltration of Nicotiana benthamiana was performed following a
reported protocol42. After infection for 48 h, the
leaves were examined by using an Olympus FV1000confocal microscope
(Japan).