Expression of a human proenkephalin A cDNA in Escherichia coli.
A 1000 base pair cDNA coding for the entire human proenkephalin A(proA) polypeptide was subcloned into the multifunctional pMPV 2911/M E. coli vector. The recombinant plasmid was found to express an approximately 30 kDa prohormone, which was recognized by a Met-Arg6-Phe2 antibody, directed against the C-terminal part of the enkephalin A prohormone. The expression of human proenkephalin A cDNA should thus permit the rapid purification of unfused recombinant enkephalin A prohormone, which itself may provide a model substrate to identify endoproteolytic processing activities.
Monstein
H J
HJ
Dept. of Clinical Chemistry, Laboratory of Molecular Biology, Copenhagen, Denmark.
D002872:Chromosome Deletion; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004745:Enkephalins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics*; D011498:Protein Precursors / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004745:Enkephalins; D011498:Protein Precursors; D011994:Recombinant Proteins; C029992:proenkephalin; C033660:preproenkephalin