HOW TO INCREASE EXPRESSION OF CLONED PROKARYOTIC GENE IN E.COLI

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dogramicrobiology
dogramicrobiology's picture
HOW TO INCREASE EXPRESSION OF CLONED PROKARYOTIC GENE IN E.COLI

hello everybody,
                                 i have cloned a 1.1 kb bacterial outer membane gene in expression vector pet22b+, i have been expressed it in BL21 cells. by inducing with IPTG (0.5mM AND 1mM final concentration. i have tried at 37 and 25 *c temp. but i could not get expression on SDSPGE . expression was only be detected by doing western blot with Anti HIS TAG ANTIBODIES. so if some one have good ideas to increase the expression then please help my clone to express significantly

waiting positively.............

Chin Fen Teo
Chin Fen Teo's picture
 Hi dogramicrobiology,

 Hi dogramicrobiology,

If your lab's budget allows, I recommend Magic Media from invitrogen. While I have no idea what kind of ingredient contributed to the "magic" factor, it does help me to increase the yield of one of the protein that i worked with comparing to the yield from IPTG induced protein expression.

Good luck.