Making Recombinant Proteins
Posted 13 May 2009 - 08:03 AM
Can anyone recomend a protocol/manual/book/journal paper that describes the process of making this recombinant protein? Any help for this newbie would be greatly appreciated!
Posted 14 May 2009 - 10:44 PM
1) you need to know the sequence of your gene.
2) Once you have the sequence, you have decide what host to use for expression (E coli, Yeast, Insect cell?)
3) Then you choose appropriate vectors (eg: pET for E coli, pGAL for yeast... )
4) You can decide if you want fusion partner, eg 6xHIS tag, FLAG tag, GST tag etc
5) Then you start doing the cloning bits and transform into your host
6) Do expression tests
Posted 15 May 2009 - 04:55 AM
Molecular Cloning: A Laboratory Manual (Sambrook)
An essential book for molecular biology labs