Construction and functional analysis of pBI105, a plant expression vector to facilitate cloning and recombinant protein purification

Binary vectors such as pBI121 are widely used for introducing genes of interest into plants via Agrobacteriun-mediated transformation method. These vectors usually have low number of unique recognition sites of restriction enzymes in their multiple cloning sites. They also lack elements such as sequences necessary for protein purification. In this article، a new plant expression vectorwith a developed multiple cloning site including recognition sites of 13 restriction enzymes، a sequence for coding 8xHis-Tag to purify recombinant proteins and the sequence of M13 reverse sequencing primer، is introduced. Construction of new vector was confirmed by PCR، digestion pattern analysis، and sequencing. The T-DNA regions of the new vector and pBI121 as a control sample were transformed to Nicotiana tabbacum L. cv. Samsun by using Agrobacterium tumefaciensstrain LBA4404. Integration of T-DNA of the vector to the genome of plantlets، which were regenerated on selective culture، was analyzed by PCR. GUS assay also confirmed expression of the transgene in transgenic seedlings. These evidences indicated that the new plant expression vector، pBI105، is efficient for plant transformation.