Expression and partial purification of human pro-opiomelanocortin in Escherichia coli.
A large portion of the human pro-opiomelanocortin (POMC) peptide corresponding to amino acid residues 59-241 has been cloned and expressed in Escherichia coli. A 1.0 kb DNA fragment encoding this peptide was cloned into the expression vectors pUC8 and pUR291. Plasmid pJMBG51 (a pUC8 recombinant) was found to direct the expression of a 24 kDa peptide. The recombinant pUR291 (pJMBG52) was shown to produce a beta-galactosidase fusion protein of 140 kDa. Western blot analysis showed that both the 24 kDa and 140 kDa peptides are recognized by antibodies raised against POMC-derived peptides. The beta-galactosidase fusion protein has been partially purified from crude E. coli cell lysates using affinity chromatography on p-amino-benzyl-1-thio-beta-D-galactopyranoside agarose.
Grewal
T S
TS
School of Animal and Microbial Sciences, Department of Biochemistry & Physiology, University of Reading, Whiteknights.
Lowry
P J
PJ
Savva
D
D
D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D007438:Introns; D010957:Plasmids; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping
D011993:Recombinant Fusion Proteins; D011333:Pro-Opiomelanocortin; D004247:DNA