Expression in Escherichia coli of chemically synthesized gene for a novel opiate peptide alpha-neo-endorphin.
Chemically synthesized alpha-neo-endorphin gene was fused to the Escherichia coli beta-galactosidase gene on the plasmid pKO13. The resulting recombinant DNA was used to transform E. coli cells. Radioimmunoassay for alpha-neo-endorphin in CNBr-treated bacterial cells showed that alpha-neo-endorphin was synthesized at approximately 5 x 10(5) molecules per single E. coli cell. One of the transformants, WA802/p alpha NE2, was used for alpha-neo-endorphin purification. From 10.9 g of wet cells, we isolated 4 mg of chemically pure and biologically active alpha-neo-endorphin.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004723:Endorphins / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D010957:Plasmids; D011498:Protein Precursors / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D004723:Endorphins; D011498:Protein Precursors; C025681:alpha-neoendorphin; D004247:DNA; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase