Genetic Construction and High-level Gene Expression in Escherichia coli of a Precursor of Salmon Calcitonin I(Biological Chemistry)

概要

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A precursor of salmon calcitonin I (SCT) was produced in Escherichia coli transformed by recombinant plasmids. A double-stranded DNA coding for SCT-Gly, preceded by ATG and followed by tandem stop codons, was constructed using a combination of chemical synthesis and enzymatic assembly. The gene for the N-terminal portion of metapyrocatechase (C23O) and its preceding ribosome binding site derived from pseudomonas putida were fused to the synthetic gene. Introduction of this fragment into E. coli resulted in the synthesis of the fusion protein at high efficientcy under the control of the tac promoter. After cleavage of the protein with cyanogen bromide, SCT-Gly was purified to homogeneity by gel filtration and by reverse phase chromatography. The structure of the peptide was confirmed by amino acid composition and sequencing analyses. The hypocalcemic actibity of the bacterial product was estimated as about one-seventh of that of SCT.
社団法人日本農芸化学会の論文
1988-11-23