membrane proteins, alkane monooxygenase (1100 bases) and alcohol
dehydrogenase (1050 bases).. The genes are from Pseudomonas aeruginosa
and I have successfully cloned them both into two vectors: pHIS1 and
pet28 (

the genes are present and they are in the correct reading frame. after induction using 1mM IPTG at 0.6 OD for 8 hrs, I am getting a similar protein pattern in both the induced and non induced. I have also tried
varying the IPTG concentration (0.1mM and 0.5mM), incubation
temperature (28 and 30 degrees celcius) and host strain (BL21 DE3
pLysS). Moreover, I have also tried 1% Triton - X treatment, liquid
nitrogen freeze-thawing and sonication of the induced samples. It has
been six months and I have not been successful in expressing both
these proteins. I would really appreciate it if anyone has any
suggestions to solve this..thank you in advance =)
Edited by heraa_daas, 26 March 2012 - 12:58 AM.