I have cloned a mutant human polb sequence in a pET28a vector. I have sequenced the clone, the sequence is in reading frame. When I try to expresses the protein in BL21 with IPTG, get no product. I have tried for several time, also in different concentration of IPTG, time period and temperature. Please help me.
Submit your paper to J Biol Methods today!
2 replies to this topic
#2
bob1
-
- Global Moderators
-
- 6,554 posts
Thelymitra pulchella
550
Excellent
Excellent
Posted 10 April 2017 - 07:53 AM
Which fractions have you solubilised and done westerns for?
#3
labtastic
-
- Active Members
-
- 205 posts
Veteran
22
Excellent
Excellent
Posted 10 April 2017 - 09:27 AM
And do know this protein should express in E.coli (from literature, previous lab members, etc)? Sometimes proteins just don't express in Ecoli.
Edited by labtastic, 10 April 2017 - 09:31 AM.
