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Applied Microbiology and Biotechnology

, Volume 31, Issue 5–6, pp 518–523 | Cite as

High-level expression of human proapolipoprotein A-I in Escherichia coli using expression plasmids containing tandemly polymerized proapolipoprotein A-I structural genes

  • Tatsurou Shibui
  • Michiru Uchida-Kamizono
  • Yasuko Takizawa
  • Jun Kondo
  • Chiharu Hiyoshi
  • Satoru Murayama
  • Yuuki Morimoto
  • Yutaka Teranishi
Applied Genetics and Regulation

Summary

A gene coding human proapolipoprotein A-I (proapo A-I) was inserted into a plasmid with a consensus ribosome binding sequence in Escherichia coli. One to three copies of this gene were tandemly inserted to construct proapo A-I expression plasmids, pMTpAI, pMT(pAI)2 and pMT(pAI)3, respectively. The cells harbouring pMT(pAI)3, could produce proapo A-I at a level of 49 μg/ml A600 and up to approx. 48% of the total cellular protein. The product was soluble in E. coli, and formed protein-lipid complexes with dimyristoyl phosphatidyl choline, which formed stacked disc structures. Analyses of the rec proapo A-I formed in the bacteria was identical to human proapo A-I except for methionine at the N-terminus.

Keywords

Escherichia Coli Methionine Choline Structural Gene Phosphatidyl Choline 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Tatsurou Shibui
    • 1
  • Michiru Uchida-Kamizono
    • 1
  • Yasuko Takizawa
    • 1
  • Jun Kondo
    • 1
  • Chiharu Hiyoshi
    • 1
  • Satoru Murayama
    • 1
  • Yuuki Morimoto
    • 1
  • Yutaka Teranishi
    • 1
  1. 1.Biosciences Laboratory, Research CenterMitsubishi Kasei CorporationYokohamaJapan

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