Internal ribosome-binding site directs expression of parathyroid hormone analogue (8-84) in Escherichia coli.
Expression of the human parathyroid hormone (PTH) gene in E. coli yielded intact PTH and PTH-(8-84). To determine if PTH-(8-84) is the result of a competing translation initiated from methionine codon-8 or degradation of the intact PTH, twelve new gene constructs with or without an internal ribosome-binding site (iRBS) in the PTH-(1-5) region were prepared via substitution with degenerate codons. Expression of constructs without iRBS produced only intact PTH. Constructs with weak iRBS, including one that resembles the cDNA sequence, yielded PTH-(8-84) as a minor product. In contrast, constructs with strong iRBS produced predominantly or exclusively this shorter analogue.
Sung
W L
WL
Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON.
Luk
C K
CK
Zahab
D M
DM
Barbier
J R
JR
Lafontaine
M
M
Willick
G E
GE
D001665:Binding Sites; D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005813:Genes, Synthetic; D006801:Humans; D009838:Oligodeoxyribonucleotides; D010281:Parathyroid Hormone / Q000235:genetics*; D010446:Peptide Fragments / Q000235:genetics*; D010957:Plasmids; D012270:Ribosomes / Q000378:metabolism*
D003062:Codon; D009838:Oligodeoxyribonucleotides; D010281:Parathyroid Hormone; D010446:Peptide Fragments; C054602:parathyroid hormone (8-84)