Farhad Shokouhifar; Nahid Abbaspour; Sahba Toosi; Nayereh Sadat Ghafarinia
Abstract
Transient expression in the plant leaf cells is a relatively fast technique to analyses regulatory sequences. To gain the approach advantages, a convenient expression vector would be needed for transient analysis of regulatory elements. In this study, to construct such a vector we used pBGi which provides ...
Read More
Transient expression in the plant leaf cells is a relatively fast technique to analyses regulatory sequences. To gain the approach advantages, a convenient expression vector would be needed for transient analysis of regulatory elements. In this study, to construct such a vector we used pBGi which provides the regulatory elements cloning site::minimal promoter as insert and pCAMBIA3301 that after removing the CaMV 35S promoter was used as backbone. The new vector, pCaBGi was constructed by replacing BamHI/SnaBI fragment (cis-acting cloning site::Minimal promoter) of pBGi with a BamHI/SnaBI fragment (CaMV 35S promoter) isolated from the pCAMBIA3301. The recombinant colonies were screened using colony PCR with and accuracy of the construction steps has been confirmed by sequencing using PSh4-F2/R specific primers. GUS assay showed that the intron containing GUS gene do not expressed agrobacterium strain GV3101 as a prokaryotic system. Injection of GV3101 harboring pCaBGi in tobacco leaves showed very low Basal expression of minimal promoter. The Sequence of pCaBGi has been submitted to gene bank using sequin software with the Accession number MG719235.
