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[wolffia]

hi there! I'm puzzled these days since I've been inducing a recombinant protein construct for certain times, but there seems no obvious induction band.
The gene constructed is a full length one ,and it's coding a transmembrane kinase,about 75kD. What I use is the pET28a system and the BL21(DE3)strain. The construct is no wrong as the sequencing result suggested. And the inserted gene is in frame. I've tried two final concentration of IPTG as 1mM and 0.4 mM, and the induction time from 1h to 20h.But there still seems no obvious band on the gel.
Could any friend tell me whether I was wrong at some place or what should be taken into account that I've missed? Thanks a lot from the bottom of my heart!