Construction of expression vector PVBG2307 by adding transcription initiation and termination elements.

Shahjahan Shabbir Ahmed; Nazeer Ahmed; Imran Ali Sani; Muhammad Naeem Shahwani; Mohammad Saeed; Sarwat Afridi; Muhammad Azam Khan; Gong Zhen Hui

Cloned elements of expression vector pB1121 carrying transcription initiation codon namely cauliflower mosaic virus section 35S and a nucleotide sequence NOS functioning transcription termination codon of cloning plasmid pB1221 were inserted into a precursor cloning plasmid pCAMBIA2300 to construct an intermediate vector termed as pVBG2307. Resultant vector pVBG2307 offers vast multiple cloning site with an adjacent imported CaMV35S promoter and transcription stop codon sequences on its left border. This vector allows controlled transformation and regulation of nucleic acids in both E. coli and Agrobacterium tumefaciens. Many cloned CaPG, orf456, ipt genes and E8 (a fruiting promoter), were amplified from cDNA libraries of sweet pepper (Capsicum annum) and tomato (Lycopersicon esculenturn) and were then transferred into vector pVBG2307. The viability of this vector was demonstrated, as it regulated CaPG, orf456, ipt and E8 genes in Escherichia coli and could be transferred into tumor inducing Agrobacterium strain EHA 105-4.