In the last we had the discussion on very fundamental concepts of how Agrobacterium affects and transfer its part of plasmid in to the genome of host plant cells? Agrobacterium is very efficiently used in genetic engineering as a vector to transfer foreign gene, we CUBists often discuss Agrobacterium in the context of Cardamine model system, where we are trying to incorporate a foreign gene in to Cardamine plant by dipping the aerial parts of the plant in to the suspension of Agrobacterium culture.
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Summary of the discussion quoted by @abhjieet Singh
The discussion started like we study about rDNA(recombinant DNA technology)in our curriculum, so how the floral dip method that is being developed can give us a context to understand our curriculum better?!
So basically Agrobacterium Tumefacien is widely used in conventional method of genetic engineering (rDNA technology)
TiPlasmid(Tumour inducing plasmid) DNA is actually used in the process. So basically a bacteria has this plasmid and when the plant get a wound or something it releases phenolic compound at that particular site. The virulent gene in the plasmid of bacterial cell gets activated because of this. And it sends the T-plasmid that contains genes producing growth regulators in the cell of the plant causing uncontrolled growth of cell so as food will be prepared for bacteria.
Conventional genetic engineering used the same mechanism but just instead of the gene producing growth regulators, scientists use their gene of interest through rDNA tech. Through restrictions enzymes and DNA ligase they replace the gene with their gene of interest and we discussed how actually does both of these enzymes work
As mentioned earlier for the activation of Virulent gene( it acts as a bridge between host cell and bacterial cell), release of phenolic compound are important but in some cases the phenolic compound isn’t secreted like in monocots. So we give some phenolic compound in the system.
We talk about all sorts of stuff and terms but do we actually understand the fundamentals of it? Like we are talking about gene transfer but do we know how does it takes place through the processes like conjugation or transformation of transduction?
In this context we discussed how and why gene transfer takes place and how conjugation or sexual reproduction help in more variations than binary fission or asexual reproduction in bacteria as it gives genetic variation that is helpful in survival. We discussed a little on vertical gene transfer (VGT) and horizontal gene transfer (HGT)
When we were discussing about rDNA tech in conventional method of genetic engineering we tried to connect it with floral dip method and some very interesting and very very important questions came up
A we discussed that virulent gene requires phenolic compound for its activation but as far as floral dip method is considered we don’t use any such compound in the suspension or in any step of it. So HOW DOES THE GENE OF INTEREST(for example kanamycin resistent gene) THAT WE WANT TO INCORPORATE IN THE PLANTS GENOME WILL REPLICATE SO AS TO ENTER THE PLANT CELL?
What is the piece of the puzzle that is Missing here? @Kiran Di @Manisha Rani Cube Ranchi Cardamine @Chitralekha Ma’am Cube @Sarita Ma’am cube @Batul Ma’am @Arunan Sir @Saida Di @Rechel cube Ranchi @Binita Cube @Shalini Lal
Then we discussed about selection of the market gene.
Why only kanamycin antibiotic and not penicilin or other antibiotic?
Mechanism or mode of action of different antibiotic etc.
Please add your points…