Question

Yay, im working on manipulating and cloning DNA for a week to insert insulin gene into E.coli BL21, should be fun. Throw down any questions you have on the topic and ill get back to ya :)

Answer 1

Look fun. What restriction enzymes are you going to use for the job? and what vector?

Answer 2

pGEX-4T-1 plasmid is the vector, the insulin gene isnt too big so it suits perfectly. Were going to use Eco R1 & Xho1, gives a good cut and the insulin gene doesnt have them in its exon

Answer 3

Wich it was me... :/
All i have is bloody patch clamp equipment... but worked quite alright for my test.

Answer 4

ah itll do, you may get slight more contamination but you can purify out

Answer 5

What is the main purpose of the project? I see the idea in having a cell creating insulin, but is there some kind of twist or are they for further study?

Answer 6

Something like genetically modify keratinocyte to produce insulin, when the gen is triggered ? Would help them with type 1-diabetes. (just thinking out loud, know you work with E.coli, but you get the point.)

Answer 7

Its mainly putting principles into practice, if we go on for thesis etc, its a good area to be familiar with, i have ideas for viruses and protection. its an area ill be relying heavily on when i get round to the project

Answer 8

I wish you luck with the project, hope you will keep me/us updated through reports or if you make a scientifically breakthrough. :)

Answer 9

Thanks, if i make an unlikely breakthrough ill let ya all know ;)