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Modification of Ti Plasmid to Construction dTi Vector for Gene Cloning
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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 beta-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 beta-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.
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