Preparation and characterization of bovine growth hormones produced in recombinant Escherichia coli.
Two analogues of bovine growth hormone (BGH) have been produced in Escherichia coli by recombinant DNA techniques. In analogue Delta-1, the N-terminal alanine residue of the full-length bovine sequence is replaced by methionine. In analogue Delta-9, which is expressed at much higher levels than is Delta-1, the full-length bovine sequence is truncated at the N-terminus by eight residues and there is a serine-for-glycine substitution in the first position of the truncated protein. Both analogues, which were characterized by isoelectric focusing (i.e.f.), polyacrylamide-gel electrophoresis in the presence of SDS (SDS/PAGE), amino acid analysis and N-terminal amino acid sequence determination using combined g.l.c.-m.s., are compared with BGH isolated from pituitaries. In contrast with pituitary-derived BGH, the recombinant-derived proteins are homogeneous on SDS/PAGE and on i.e.f. In a radioimmunoassay, a radioreceptor assay and a bioassay in vivo (rat tibia), Delta-9 BGH showed very similar characteristics to the pituitary-derived hormone. Similar results have also been obtained with the Delta-1 analogue.
Wingfield
P T
PT
Biogen S.A., Geneva, Switzerland.
Graber
P
P
Buell
G
G
Rose
K
K
Simona
M G
MG
Burleigh
B D
BD
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D004220:Disulfides / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D013006:Growth Hormone / Q000235:genetics / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D011863:Radioimmunoassay; D011995:Recombination, Genetic; D013438:Sulfhydryl Compounds / Q000032:analysis
D000596:Amino Acids; D004220:Disulfides; D013438:Sulfhydryl Compounds; D013006:Growth Hormone