Brief note about Ti-plasmid

Brief note about  Ti plasmid

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Ti plasmid is a large mega plasmid conjugative plasmid of ~200kb. pTi is lost when Agrobacterium is grown above 28oC, such cured bacteria do not induce crown galls that is, they become a virulent. pTi and pRi, although do not share sequence homology but are unique in following respects:- a) They contain some genes, which are located within their T-DNA which has regulatory sequences recognized by plant cells, while their remaining genes have prokaryotic regulatory sequences. As a result, the former are expressed only in plant cells but not in the Agrobacterium, while the latter are only expressed in the bacterium. b) These plasmids naturally transfer a part of their DNA, called as T-DNA, into host plant cells. The T-DNA usually contains following important functional regions.
1. T-DNA contains oncogenes and opine synthesis genes and is transferred into host plant.
2. Vir region which regulates the transfer of T-DNA
3. Opine catabolism genes for utilization of opines.
4. Origin of replication for propagation in Agrobacterium. The T-DNA contains a 24bp direct repeat border sequence and contains the genes necessary for tumor / possess gene for auxin and cytokinin biosynthesis. All the genes present in TDNA have eukaryotic regulatory sequences. As a result, these genes are expressed only in plant cells but never express in Agrobacterium. The vir region mediates the transfer of TDNA into plant genomes and hence is essential for virulence. The genes of vir region are not transferred but induce the transfer of T-DNA. Also, the genes present in T-DNA are not responsible for its transfer, but the 24 bp direct repeat at both the left and right ends of TDNA is essential for the transfer. The exact mechanism of transfer of T-DNA is not known clearly known but is brought by the vir region. The phenols produced by wounded plant tissue initiates the transfer process. The T-DNA is transferred into the plant cells as single stranded DNA, which increases the efficiency of its transformation. But, as soon as it enters into the plant cell, it is immediately converted into a double stranded form. This form integrates at random sites in the host plant genome by a phenomenon called illegitimate recombination, which are due to sequence of homology in short segments of the host DNA. This integration is usually in low copy numbers. Few vectors are derived from pTi (wild type) due to some problems posed by wild type plasmid eg. The presence of oncogenes causes a disorganized growth, their large size and lack of cloning sites within the T-DNA, which are needed for the insertion of DNA segments that has to be cloned.