NO EXPRESSION IN LARGE SCALE - (Dec/21/2007 )

HI THERE
I AM TRYING TO EXPRESS A PLANT GENE IN E-COLI EXPRESSION SYSTEM. I HAVE USED pGEX EXPRESSION VECTOR INTO BL21 BUT I GET ALITLE PROTEIN IN SMLL SCALE AND NO EXPRESSION IN LARGE SCALE. THEN I CHANGED THE VECTOR TO pETM-41 BUT STILL THERESE THE SAME PROBLEM.
I WOULD BE GLAD IF SOMEBODY HELP ME IN THIS CASE.
THANK YOU VERY MUCH.

BEST,

For how long did you induce the production of the protein with IPTG? And at witch concentration?

Have you checked the insoluble fraction of the bacteria? The overexpression of a protein in bacteria often forms inclusion bodies, witch are insoluble.

i agree

What is ur small and large scale volume?