Recent replies to "protein expression in bacteria"

Reply from Maria Noutsou

Maria Noutsou — Fri, 11 Apr 2008 19:16:38 -0000

Hello João,

I don’t know if you found the answer to your question already, but I would like to suggest you a few things.
Use pET bacterial expression vectors. You can find a variety. The basic differences are on the location of purification tags and Multicloning regions. These vectors have a selective marker (antibiotic) and also they provide you a strong promoter upstream of your gene. This promoter is able to be regulated by IPTG. So, you will grow your cells until OD600 0,6-0,8 and then you need to add 0,4-0,8 mM IPTG in order to induce the expression of your protein.
You don’t want to have expression from the begining because it might interfere with the growing of your bacteria. In addition, I would suggest to use bacteria strains which are desinged for protein expression like Rosetta2 or BL21 pLysS.

I hope this will help,

Maria

Reply from Farasha Almadani

Farasha Almadani — Thu, 27 Mar 2008 21:28:23 -0000

Hi Joao

I dont know why you want to use a mammalian vector for this purpose. However, from my own work I use pET21b from Novagen it is quite good expression system or you can use pQE expression system from Qiagen . Both are for bacterial expression of recombinant proteins.

Hope this helps

Regards
Farasha

Reply from Lisa Ali

Lisa Ali — Thu, 20 Mar 2008 22:14:26 -0000

I have no idea what you are talking about..but it sounds very interesting.