Not being a plant biologist I can't speak directly about integration into the apple genome but I can give you some general input about genomic integration. First of all you will some sort of selectable marker in your input DNA-this is usually a drug resistance gene, or a gene necessary for biosynthesis of an essential nutrient. You will probably have to search through the literature to find what types of markers are being used in apples. Then your going to have to identify a target site that you wish to integrate into so that you can design your input DNA to have homology at the ends to the target site. So a basic DNA construct for integration would look like this: DNA sequence homologous to the target site followed by your gene of interest followed by a selectable marker (both of these will need appropriate promoters) followed by DNA sequence homologous to the target site. Some constructs also have sequences flanking the selectable marker so that it can be excised at a later date.
Some organisms can be very effeciently transformed and integrate DNA into the chromosome (i.e. yeast- Saccharomyces), others such as E. coli will degrade incoming linear DNA so you need to coexpress a recombinase. Not sure what is required for apples-I again would search the literature.
I would caution you to think twice about fooling around with agriculturally important species such as apples or other food crops for a couple of reasons; first of all there is quite a bit of controversy and regulation (depending on where you live) concerning GMO crops. If you start messing around in your basement you might attract a lot of attention that you probably don't want and may have some government agency come shut down your home lab for violating some obscure regulation. Secondly I think you have to think seriously about containment and making sure that what ever you create doesn't escape. It's unlikely, but possible, that what you create could spread through apple populations with really unknown consequences. Before proceeding you should do a very through risk assesment and consult with plant biologists from the apple community.
Some may shrug at my caution but I think that each of us owes it to the larger DIY-Bio community to be extremly circumspect about what we are doing in our labs. Theres plenty of suspision and apprehension about DIY Bio and all it would really take is some inadverdent release or accident to raise the barriers to these kinds of activities.
My two cents!
On Wednesday, May 16, 2012 6:36:05 AM UTC-4, Mega wrote:
Ah.... Read more about it. is it true that quite every plasmid can be integratet into the Chromosome when you keep it very long in the antibiotic media? Also in animal cells? The chromosome is big, so there probably will be a sequence for homologous recombination?--
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