Expression of the cDNA for alpha 1-acid glycoprotein, a rat plasma protein, in Escherichia coli.
Polyadenylated RNA was isolated from acute-phase liver and transcribed into cDNA. After homopolymer tailing this was cloned into the PstI site of pBR322 which was used to transform E. coli RR1 cells. Clones containing cDNA corresponding to acute-phase proteins were identified by differential hybridization with 32P-labelled normal and acute-phase cDNA. A clone synthesizing about 10 ng of alpha 1-acid glycoprotein per E. coli colony was detected using a solid-phase based immunochemical procedure. The identification of the clone was verified by competition assay with authentic alpha 1-acid glycoprotein isolated from serum and by restriction analysis of the cDNA inserted into pBR322.
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D007158:Immunologic Techniques; D008099:Liver / Q000032:analysis; D009693:Nucleic Acid Hybridization; D009961:Orosomucoid / Q000096:biosynthesis / Q000235:genetics*; D011061:Poly A / Q000235:genetics; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger; D051381:Rats
D009961:Orosomucoid; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004247:DNA; D004262:DNA Restriction Enzymes