RT Journal T1 NUCLEAR LOCALIZATION OF ARABIDOPSISĖˆS HEAT SHOCK FACTOR HSFA1D USING BIMOLECULAR FLORESCENCE COMPLEMENTATION (BIFC) SYSTEM A1 Z. Shah A1 G S. Ali A1 A. El-Sayed A1 M. Hussain A1 H. Shah A1 M. Z. Ahmad A1 N. Ahmad 1and A. Iqbal JF Journal of Animal and Plant Sciences JO JAPS SN 1018-7081 VO 31 IS 4 SP 1160 OP 1166 YR 2020 FD 2020/12/18 DO DOI https://doi.org/10.36899/JAPS.2021.4.0313 AB

Response to thermal stress in plants is mostly regulated by heat shock factors (hsfs). Among hsfs, HsfA1d is one of the key players in protecting plants against heat stress. Subcellular localization of gene is prerequisite for its characterization. Bimolecular Fluorescent Complementation (BiFC) provides the most advance and reliable tool to visualize protein-protein interactions and to explore their location in living system. Objective of the present study was to accomplish sub cellular localization of HsfA1d by using BiFC. HsfA1d was cloned in-frame with complementary halves of yellow florescent proteins (YFP) using gateway cloning system. The BiFC constructs along with positive control were introduced into plant cells through agro-infiltration. The non-florescent fragments of YFP tagged with HsfA1d produced no signal when they were infiltrated alone in tobacco leaves. The N and C termini of YFP fused with HsfA1d resulted in florescence upon co-expression in tobacco leaf epidermal cells. The intact YFP tagged with HsfA1d, after DAPI staining, was observed under 3 different laser beams of YFP, DAPI and merged. The same florescent signals detected under different channels of laser scanning confocal microscope confirmed the nuclear localization of HsfA1d. It is concluded, that HsfA1d is localized in the nucleus of cell.

K1 HsfA1d, subcellular localization, yellow florescent proteins, gateway cloning, BiFC PB Pakistan Agricultural Scientists Forum LK https://thejaps.org.pk/AbstractView.aspx?mid=Biot-20-0004