2 replies to this topic

[anandraj79]

Hi,

I am struggling with protein expression, my protein nature is toxic to host cell. i have tried pEXP5-CT/TOPO expression vector system but didn`t work out.

Suggestion from all researcher are mostly welcome.


regards,


Anand

[Dr Teeth]

anandraj79, on Sep 25 2009, 08:04 AM, said:

Hi,

I am struggling with protein expression, my protein nature is toxic to host cell. i have tried pEXP5-CT/TOPO expression vector system but didn`t work out.

Suggestion from all researcher are mostly welcome.


regards,


Anand

If your protein expression is toxic to the target cells, why would you use pEXP5?  This vector is designed for high level expression.  You want something with reduced expression or better yet, an inducible system, such as tet-on, where your protein would not be expressed unless the target cells were treated with Doxycyline.

[andy.dods6]

Dr Teeth, on Sep 25 2009, 08:54 AM, said:

anandraj79, on Sep 25 2009, 08:04 AM, said:

Hi,

I am struggling with protein expression, my protein nature is toxic to host cell. i have tried pEXP5-CT/TOPO expression vector system but didn`t work out.

Suggestion from all researcher are mostly welcome.


regards,


Anand

If your protein expression is toxic to the target cells, why would you use pEXP5?  This vector is designed for high level expression.  You want something with reduced expression or better yet, an inducible system, such as tet-on, where your protein would not be expressed unless the target cells were treated with Doxycyline.

Just a note to this point, if you were going to produce a low expressing vector, try sticking a Z-tag on it.  This enables very good capture of your protein, allowing maximal enrichment.