How is a gene isolated and inserted in genetic engineering?

Question

I heard there were few different techniques involved and I am not sure if there are actually true. Can someone enlighten me on the subject? Is there just one technique that is used? If so, how is it done?

Answer 1

There are many, many techniques for this. Gene isolation can be done by screening a cDNA library, a genomic library, an expression library, differential display, among others. Basically you are looking to identify a gene by function, or you already know part or all of it's sequence so you can isolate it by hybridization.

For insertion using genetic engineering, you can use a plasmid vector and use bacterial infection, or a viral vector and use transfection, or you can inejct the DNA directly into the cell as well. Even artificial chromosomes are on the verge or being used.

Those are the options. The details are fairly lengthy though if you are looking for protocol. Is that what you want?

Answer 2

I was reading that the have just successfully used the first nano-technology to insert something into a cell. I would think they will eventually be able to program nono-machines to track down a cell with a corrupted DNA strand and insert a repaired one, removing the damaged DNA and expelling it from the body.

With stem cells they insert the DNA with a needle, or something similar, I have seen the images on a show here and there. The thing with doing it that way is it is damaging to the cell. Nano-technology finds a way to go through the cell wall with a way of forced osmosis, leaving the cell membrane intact.

I also read that they are now able to build complex proteins artificially and are planning to insert them into damaged cells, for instance with Alzheimer’s, there is a broken protein that infects the building of other proteins. They could perhaps use this artificial protein to break the chain, so to speak.

Answer 3

restriction enzymes