Expression of an Artificial Gene Coding for a Human Secretin Precursor-Like Peptide in Escherichia coli

概要

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A gene coding for human secretin elongated at the C-terminus by Gly-Lys-Arg was chemically synthesized and fused to the downstream of an artifical gene encoding [Leu^<14>]human growth hormone (hGH) which had a leucine residue instead of methionine at position 14 in the wild type hGH protein. The fusion gene was efficiently expressed under the control of the Escherichia coli trp promoter including the consensus Shine-Dalgarno sequence. After the fusion protein was cleaved in two parts with cyanogen bromide, the desired secretin-Gly-Lys-Arg (SecGKR) was finally purified to homogeneity using high-performance liquid chromatography, yielding 11 mg protein from 16 g of wet cells. The biological activities of secretin-Gly-Lys (SecGK) and secretin-Gly (SecG) obtained by digestion of SecGKR with carboxypeptidase B were compared with SecGKR and the mature human secretin. The activities of SecGKR and SecGK were 25-30% higher than that of the mature human secretin amidated at the carboxy-terminal amino acid, but that of SecG was 25% lower.
社団法人日本生物工学会の論文
1991-09-25