I need to produce Oxalate decarboxylase enzyme as a recombinant enzyme. I choosed my source organisim named bacillus subtilis 168 designed my primers (with doubt) and I want to express in pET-SUMO vector. Are there anyone who tried this vector to produce protein for enzymatic reaction ? OR Can you please suggest me a vector system to produce protein for enzymatic reaction ?
I suggest you follow published procedures.
Anand et al., Biochemistry 2002, 41, 7659-7669
Production os recombinant protein may not be a straight walk as it looks on the surface. Sometimes they are many huddles to jump. I am not sure from the information you have provided if you have already obtained the gene of interest. However, if you want to amplify it directly from your Bacillus strain, then try the CODEHOP strategy. Secondly, use an expression system that allows you to explore the three alternative reading frames and choose one that will produce the protein of correct size as your. Thanks. Good Luck.
Joseph
Thanks a lot for your answers. @adam B Shapiro ; I have already read this puplition but it is a bit "old" there are new expression system and I want to choose them.
@joseph Hawumba; I could not understand CODEHOP strategy :(
Lorriane clark; I have pET-SUMO expression system and so I also have DE3 competent cells
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