Purification and Characterization of the Product of Chemically Synthesized Human Growth Hormone Gene Expression in Escherichia coli(Biological,Chemical)

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概要

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• The efficient purification and characterization of the product of chemically synthesized human growth hormone (hGH) gene expressed in Escherichia coli are described. The product was purified from the cell lysates. of the E. coli by means. of ammonium sulfate precipitation, DE-52 chromatography, chromatofocusing chromatography and Ultrogel AcA 54 chromatography. The purified hGH gene product was homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, non-denaturing polyacrylamide gel electrophoresis, gel electrofocusing and high-performance liquid chromatography (HPLC). The purified product and an authentic methionyl hGH (m-hGH) showed identical behavior in these systems. The structural features of the purified product were examined by means of amino acid composition analysis, NH_2-terminal sequence analysis and tryptic peptide mapping. The experimental values of the amino acid composition of the purified product were in agreement with the theoretical values for m-hGH. Its NH_2-terminal sequence (39 amino acid residues) was identical with that of the published sequence of hGH, except for an additional amino-terminal methionine residue immediately preceding phenylalanine at residue 1. The elution profile of the tryptic peptides of the purified product on HPLC was identical with that of authentic m-hGH. These elution profiles were nearly identical with that of a pituitaryderived hGH, with the exception of one peak due to NH_2-terminal peptide. On the basis of these results, the purified product was identified as m-hGH.

• 社団法人日本薬学会の論文
• 1987-10-25

著者

• 内田 恵理子

  国立医薬品食品衛生研究所遺伝子細胞医薬部

• 池原 森男

  Faculty of Pharmaceutical Sciences, Osaka University(Present address)

• 早川 尭夫

  国立医薬品食品衛生研究所

• 大塚 栄子

  Faculty of Pharmaceutical Sciences, Osaka University

• 内田 恵理子

  国立衛研

• 大塚 栄子

  Faculty Of Pharmaceutical Sciences Hokkaido University

• 押沢 正

  National Institute of Hygienic Sciences

• 内田 恵理子

  National Institute of Hygienic Sciences

• 山羽 力

  National Institute of Hygienic Sciences

• 寺尾 允男

  National Institute of Hygienic Sciences

• 早川 堯夫

  国立医薬品食品衛生研究所生物薬品部

• 寺尾 允男

  国立衛生試験所

• 寺尾 允男

  National Institute Of Health Sciences

• 新見 伸吾

  国立衛試・生物

• 早川 尭夫

  Division Of Biological Chemistry And Reference Standards National Institute Of Hygienic Sciences

• Uchida Eriko

  Human Genome Center Institute Of Medical Science University Of Tokyo

• 池原 森男

  Faculty Of Pharmaceutical Sciences Osaka University

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