Please Help (pMAL)

wsa wsa at st-andrews.ac.uk
Thu Aug 1 04:34:47 EST 1996


I've cloned the gene coding for my protein into pMAL-p2 at the EcoRI 
and XbaI sites but I am having problems expressing the fusion protein.
The protein itself is not toxic as I have previously expressed it in a 
pRSET vector except it was insoluble. I have tried gowing the cells in 
L-Broth with and without glucose and inducing with IPTG for 3 hours.
So far I have tried to express the fusion protein in JM105, XL1-Blue 
and BL21(DE3) cells without any success. I've tried expressing pMAL-p2 
without an insert and this failed to produce any new bands on a gel or 
blot. My antibody against my fails to pick up any bands other than the 
control and the New England Biolabs MBP antibody only detects a band 
around 50,000 Da. 

If anyone has any ideas I would really appreciate them as I am running 
out of time on this project. Thanks in advance

W.Stuart Annan



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