RNA structural elements for expression in Escherichia coli

FEBS Letters

Courtney, Michael¹  Tessier, Luc-Henri¹  Sauvageot, Martine¹  Jallat, Sophie¹  Crystal, Ronald G.² 

[1] TRANSGENE SA, 11 Rue de Molsheim, 67 Strasbourg, France;National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20205, USA

关键词: recombinant DNA Gene fusion Point mutation λplasmid vector Gene cloning Emphysema;    α1AT;    α1-antitrypsin;    Ap;    ampicillin;    Δ;    deletion;    IFN;    interferon;    IHF;    integration host factor;    LB;    Luria broth;    PA;    polyacrylamide;    PAGE;    PA gel electrophoresis;    PBS;    phosphate-buffered saline;    RBS;    ribosome-binding site;    RID;    radial immune diffusion;    TE buffer;    2 mM Tris-HCl pH 8;    10 mM EDTA;   

DOI  :  10.1016/0014-5793(86)81014-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Analysis of a series of λ cII::α₁-antitrypsin (α₁AT) gene fusions of different sizes showed that increased α₁AT expression correlated with the stabilisation of a particular computer-predicted RNA secondary structure. Moreover, significant synthesis of unfused α₁AT was achieved by reconstruction of this conformation to permit interaction between the upstream region of the ribosome-binding site and the first part of the α₁ AT coding sequence. This high-level expression was dependent upon certain silent point mutations in the coding sequence, indicating that RNA primary and secondary structure determinants can operate in concert to dictate the efficiency of protein synthesis.

【 授权许可】

Unknown