High-level expression of binase in Escherichia coli

A. A. Schulga*, A. L. Okorokov, K. I. Panov, F. T. Kurbanov, B. K. Chernov, K. G. Skryabin, M. P. Kirpichnikov

*Corresponding author for this work

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9 Citations (Scopus)

Abstract

We have reported previously about the cloning of the binase gene in E. coli. In this work, using an original approach named 'homolog gene recombination' method (HGR), vectors for binase expression in E. coli have been constructed. Transcription of the binase gene have been directed through either tac-promoter or P(R)-promoter of bacteriophage λ under the control of temperature-sensitive C1857 repressor. The last promoter gave the maximum yield of binase, up to 100 mg of protein per litre of heat-induced bacterial culture. The location of the transcription terminator at the 3' terminus of the binase gene raised the expression approximately two times more. A chromatographic method have been developed and applied for the control of binase accumulation in growth medium without measuring the ribonuclease activity.

Original languageEnglish
Pages (from-to)453-463
Number of pages11
JournalMolekulyarnaya Biologiya
Volume28
Issue number2
Publication statusPublished - 01 Jan 1994
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)

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    Schulga, A. A., Okorokov, A. L., Panov, K. I., Kurbanov, F. T., Chernov, B. K., Skryabin, K. G., & Kirpichnikov, M. P. (1994). High-level expression of binase in Escherichia coli. Molekulyarnaya Biologiya, 28(2), 453-463.