Recombinant DNA tech (for insulin)

[Doze]

Hello Community,

There is an assignment I have to do, and I chose to do research on: The Biotech of producing Human products.

I chose the production of Insulin through bacterial cells, since it's the most common, and there is something I am not understanding about the process of it though.

1)I found that Ecor1 and Hind111 cleave the Insulin protein since there is an A and B chain (is this correct?), which Restriction Enzyme cleaves which? And are both these chains needed?

2)Before the gene sequence coding for the A and B chain of Insulin from the human is transfered into the vector, does it have to be converted into mRNA first? or is it just cleaved by Ecor1 and Hind III and directly trasnferred to the vector? Or is a template made and copied from the original DNA BEFORE it's cleaved?

It's probably rather straight forward, but I'm not really seeing it. Help would be much appreciated.


Thank you.

[Cat]

Read http://www.littletree.com.au/dna.htm and then ask if you don't understand something.

[JackBean]

first of all, it's EcoRI and HindIII (the 1 are in roman style, i.e. capital i). Second, EcoRI and HindIII do not cleave protein, they are endonucleases, so they cleave nucleic acids in middle of the strain