Production of plasmids giving high expression of recombinant DNA‐derived ovine growth hormone variants in Escherichia coli

FEBS Letters

Wallis, O.Caryl¹  Wallis, Michael¹ 

[1] Biochemistry Laboratory, School of Biological Sciences, University of Sussex, Falmer, Brighton BN1 9QG, England

关键词: Growth hormone;    Growth hormone variant;    cDNA;    Expression;    Sequence;    (Ovine;    Escherichia coli);    GH;    growth hormone (oGH;    ovine GH;    bGH;    bovine GH);    IPTG;    isopropyl β-D-thiogalactopyranoside;   

DOI  :  10.1016/0014-5793(89)80758-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A method for the production of plasmids giving different levels of expression of ovine growth hormone (oGH) variants in E. coli is described. The cDNA sequence coding for mature oGH was inserted into the multiple cloning site of plasmid pUC8 and random deletions were then introduced 3′ to the initiation codon. Clones producing GH (with varying N-terminal extensions) were identified by immunological screening. Levels of expression of GH-related protein, measured by immunoassay or on SDS-polyacrylamide gels, varied from over 20% to less than 0.05% of total cell protein. The coding sequence of plasmid pOGHe101, giving very high expression of variant oGH1, was determined.

【 授权许可】

Unknown