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A prototype stable RNA identification cassette for monitoring plasmids of genetically engineered microorganismsA prototype stable RNA identification cassette for monitoring genetically engineered plasmids carried by strains of Escherichia coli has been developed. The cassette consists of a Vibrio proteolyticus 5S ribosomal RNA (rRNA) gene surrounded by promoters and terminators from the rrnB operon of Escherischia coli. The identifier RNA is expressed and successfully processed so that approximately 30% of the 5S rRNA isolated from either whole cells or 70S ribosomes is of the V. proteolyticus type. Cells carrying the identifier are readily detectable by hybridization. Accurate measurements show that the identification cassette has little effect on fitness compared to a strain containing an analogous plasmid carrying wild type E. coli 5S rRNA, and the V. proteolyticus 5S rRNA gene is not inactivated after prolonged growth. These results demonstrate the feasibility of developing small standardized identification cassettes that can utilize already existing highly sensitive rRNA detection methods. Cassettes of this type could in principle be incorporated into either the engineered regions of recombinant plasmids or their hosts.
Document ID
20040089673
Acquisition Source
Headquarters
Document Type
Reprint (Version printed in journal)
Authors
Hedenstierna, K. O.
(University of Houston TX 77204-5934, United States)
Lee, Y. H.
Yang, Y.
Fox, G. E.
Date Acquired
August 21, 2013
Publication Date
January 1, 1993
Publication Information
Publication: Systematic and applied microbiology
Volume: 16
ISSN: 0723-2020
Subject Category
Exobiology
Funding Number(s)
CONTRACT_GRANT: NAGW-2108
CONTRACT_GRANT: D86-61
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Exobiology

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