[Arabidopsis] request for info on gene cloning

md hussain via arab-gen%40net.bio.net (by md_hussain_2000 from yahoo.com)
Sun Nov 25 01:20:40 EST 2007


  Dear members,
   
  Good morning.I have the following confusions or doubts. I request you kindly clarify my doubts.
   
  1. When we are doing doing gene cloning for protein expression, why to clone a gene first in pBluescript vector and then subclone to Pet Vectors? Why don’t we directly clone the gene in pET vectors?
   
   
  2. In a gene cloning for protein expression studies, how to select starting methionine, that means when I have a methionine code in my gene and in expression vector also, then how to select the starting methionine?
   
  Could you please explain aout the inframe.And any sites, books for this?
  Thanking you in advance for your help.
  High regards,
  Hussain. 

       
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