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Studies on the plasmid instability in Corynebacteria

S Na1, T Shen, P Jia

  • 1Institute of Microbiology, Chinese Academy of Sciences, Beijing.

Chinese Journal of Biotechnology
|January 1, 1994
PubMed
Summary

Researchers constructed a shuttle vector, pNAR4, for E. coli and Coryneform bacteria. Plasmid instability was observed in Corynebacterium strains, potentially due to supercoil DNA and recombination.

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Area of Science:

  • Molecular Biology
  • Microbial Genetics

Background:

  • Shuttle vectors are crucial for genetic manipulation in diverse bacterial hosts.
  • Understanding plasmid stability is essential for efficient strain engineering in Corynebacterium.

Purpose of the Study:

  • To construct and characterize the recombinant shuttle vector pNAR4.
  • To investigate the instability of pNAR4 in various Corynebacterium strains.
  • To elucidate the molecular mechanisms underlying plasmid instability.

Main Methods:

  • Construction of recombinant plasmid pNAR4 by ligating DNA fragments.
  • Transformation of pNAR4 into different Corynebacterium strains (C. crenatum B9, C. glutamicum 10147, C. crenatum T6-13).
  • Analysis of plasmid DNA integrity and characterization using restriction digestion and DNA hybridization.

Main Results:

  • Plasmid pNAR4 exhibited instability, including segregational instability in C. crenatum B9 and structural instability in C. glutamicum 10147 and C. crenatum T6-13.
  • Transformation of pNAT65 into C. glutamicum 10147 resulted in altered plasmids resembling pXZ10145.
  • DNA hybridization revealed homology between supercoil DNA in C. glutamicum 10147 and pXZ10145, suggesting a link to instability.

Conclusions:

  • The observed plasmid instability in Corynebacterium strains is associated with the presence of supercoil DNA.
  • Inter- or intra-molecular recombination is proposed as the underlying mechanism for plasmid instability phenomena.

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