Originally Posted by monica ngini
this is an assgniment from my leturer at KEMU university;
question 5.suppose you are given a task of constructing a lasmid expression vector suitable for moleculaar cloning in an organism of pharmacutical industurial interest.for example production of insulin in E coli ,list the seps you would use to develop such a plasmid
I assume you mean, plasmid as opposed to lasmid.
To give you a brief over view of what your answer should include is the following;
You would need a strain of bacteria to work with, lets just say for example you work with DH(Alpha)5 strain of E.Coli. You want to construct a vector, that will code for insulin. Through the use of restriction enzymes, you will cut out a part of the plasmid. This will be the area where you "insert" your vector by ligation. After ligation, you should have a plasmid vector that will produce human insulin.
To actually get the desired vector from a dna sequence, you can run gel electrophoresis and use restriction enzymes to cut desired areas. With UV light, and stained gel you can cut out the desired fragment. You can then proceed with a two step ligation, or three step or what ever the case may be.