Internal ribosome-binding site directs expression of parathyroid hormone analogue (8-84) in Escherichia coli |
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Authors: | W L Sung C K Luk D M Zahab J R Barbier M Lafontaine G E Willick |
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Institution: | Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON. |
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Abstract: | 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. |
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