Modification of Ti Plasmid to Construction dTi Vector for Gene Cloning

The extracted DNA of Agrobacterium tumefaciens isolates were subjected to PCR for amplifying 16S rRNA and for amplifying T-DNA fragment then subjected to gel electrophoresis. The individual band of the 16S rRNA gene was characterized by 1479bp and of the T-DNA fragment by 1200bp. The products were comparison with the standard molecular DNA ladder (1200 and 1500bp). The purified β-lactamase gene was cut from PGLO plasmid ligated by use T4DNA ligase enzyme with the Ti plasmid which disarmed T-DNA by using the same restriction enzyme. The result was indicated by using the E.coli K12 for carries the Ti plasmid vector which contain a β-Lactamase gene when put the antibiotic ampicillin in different consternation into the LB MHA plate only the colonies which that have picked up exogenous DNA (dTi plasmid DNA ) can grow that is mean it become resistance to ampicillin by using dTi vector.