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 carrie...
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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