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

Abstract


Cloned elements of expression vector pBI121 carrying transcription initiation codon namelycauliflower mosaic virus section 35S and a nucleotide sequence NOS functioning transcription termination codon of cloning plasmid pBI221 were inserted into a precursor cloning plasmidpCAMBIA2300 to construct an intermediate vector termed as pVBG2307. Resultant vector pVBG2307 offers vast multiple cloning site with an adjacent imported CaMV35S promoter andtranscription 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 clonedCaPG, orf456, ipt genes and E8 (a fruiting promoter), were amplified from cDNA libraries of sweet pepper (Capsicum annum) and tomato (Lycopersicon esculentum) and were thentransferred 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 inducingAgrobacterium strain EHA105-4.

Keywords


pBI121, pBI221, pCAMBIA2300; pVBG2307; CaPG gene; E8 promoter

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