Construction of the plasmid-free strain for human growth hormone production

A. A. Schulga, P. V. Mechev, M. P. Kirpichnikov, K. G. Skryabin, S. M. Deyev

Research output: Contribution to journalArticle

Abstract

The E. coli strain, overproducing human growth hormone (hGH) was made by integration of the hGH gene under the control of T7 promoter into the chromosomal LacZ gene of BL21(DE3) via lambda Red recombineering. The strain gave higher productivity (50 mg·L−1·OD550 −1) and better growth characteristics than the corresponding strain in which the same hGH expression cassette was placed in a plasmid. The protein produced by the plasmid-free strain was purified and characterized to be hGH. The results demonstrates that a plasmid-free recombinant strain having a single-copy gene expression cassette in the chromosome could provide better gene activity regulation, higher productivity, superior growth characteristics, as well as more stringent control of the gene sequence invariance than a plasmid-based strain.

Original languageEnglish
Pages (from-to)148-153
Number of pages6
JournalBiochimie
Volume128-129
DOIs
Publication statusPublished - 1 Sep 2016
Externally publishedYes

Keywords

  • Human growth hormone
  • Lambda Red recombineering
  • Plasmid-free
  • Somatotropin

ASJC Scopus subject areas

  • Biochemistry

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  • Cite this

    Schulga, A. A., Mechev, P. V., Kirpichnikov, M. P., Skryabin, K. G., & Deyev, S. M. (2016). Construction of the plasmid-free strain for human growth hormone production. Biochimie, 128-129, 148-153. https://doi.org/10.1016/j.biochi.2016.08.007