Interplay of SOS induction, recombinant gene expression, and multimerization of plasmid vectors in Escherichia coli

Jeewon Lee, Hyung Cheol Kim, Sung W. Kim, Seung Wook Kim, Suk I. Hong, Young Hoon Park

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Using pBR322- and pUC-derived plasmid vectors, a homologous (Escherichia coli native esterase) and three heterologous proteins (human interleukin-2, human interleukin-6, and Zymomonas levansucrase) were synthesized in E. coli lC2015(recA::lacZ) and GY4786 (sfiA::lacZ) strains. Via time-course measurement of β-galactosidase activity in each recombinant culture, the SOS induction was estimated in detail and the results were systematically compared. In recombinant E. coli, the SOS response did not happen either with the recombinant insert-negative plasmid backbone alone or the expression vectors containing the homologous gene. Irrespective of gene expression level and toxic activity of synthesized foreign proteins, the SOS response was induced only when the heterologous genes were expressed using a particular plasmid vector, indicating strong dependence on the recombinant gene clone and the selection of a plasmid vector system. It is suggested that in recombinant E. coli the SOS response (i.e., activation of recA expression and initial sfiA expression) may be related neither to metabolic burden nor toxic cellular event(s) by synthesized heterologous protein, but may be provoked by foreign gene-specific interaction between a foreign gene and a plasmid vector. Unlike in E. coli XL1-blue(recA-) strains used, all expression vectors encoding each of the three heterologous proteins were multimerized in E. coli lC2015 strains in the course of cultivation, whereas the expression vectors containing the homologous gene never formed the plasmid multimers. The extent of multimerization was also dependent on a foreign gene insert in the expression vector. As a dominant effect of the SOS induction, recombinant plasmid vectors used for heterologous protein expression appear to significantly form various multimers in the recA+ E. coli host.

Original languageEnglish
Pages (from-to)84-92
Number of pages9
JournalBiotechnology and Bioengineering
Volume80
Issue number1
DOIs
Publication statusPublished - 2002 Oct 5

Fingerprint

Gene expression
Escherichia coli
Plasmids
Gene Expression
Genes
Proteins
Poisons
levansucrase
Zymomonas
Galactosidases
Son of Sevenless Proteins
Esterases
Interleukin-6
Time measurement
Clone Cells
Chemical activation

Keywords

  • Foreign gene-specific interaction
  • Plasmid multimers
  • SOS response

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

Cite this

Interplay of SOS induction, recombinant gene expression, and multimerization of plasmid vectors in Escherichia coli. / Lee, Jeewon; Kim, Hyung Cheol; Kim, Sung W.; Kim, Seung Wook; Hong, Suk I.; Park, Young Hoon.

In: Biotechnology and Bioengineering, Vol. 80, No. 1, 05.10.2002, p. 84-92.

Research output: Contribution to journalArticle

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