NASA Logo

NTRS

NTRS - NASA Technical Reports Server

Back to Results
A novel approach for monitoring genetically engineered microorganisms by using artificial, stable RNAsFurther improvements in technology for efficient monitoring of genetically engineered microorganisms (GEMs) in the environment are needed. Technology for monitoring rRNA is well established but has not generally been applicable to GEMs because of the lack of unique rRNA target sequences. In the work described herein, it is demonstrated that a deletion mutant of a plasmid-borne Vibrio proteolyticus 5S rRNA gene continues to accumulate to high levels in Escherichia coli although it is no longer incorporated into 70S ribosomes. This deletion construct was subsequently modified by mutagenesis to create a unique recognition site for the restriction endonuclease BstEII, into which new sequences could be readily inserted. Finally, a novel 17-nucleotide identifier sequence from Pennisetum purpureum was embedded into the construct to create an RNA identification cassette. The artificial identifier RNA, expressed from this cassette in vivo, accumulated in E. coli to levels comparable to those of wild-type 5S rRNA without being seriously detrimental to cell survival in laboratory experiments and without entering the ribosomes. These results demonstrate that artificial, stable RNAs containing sequence segments remarkably different from those present in any known rRNA can be designed and that neither the deleted sequence segment nor ribosome incorporation is essential for accumulation of an RNA product.
Document ID
20050000140
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Pitulle, C.
(University of Houston Texas 77204-5934, United States)
Hedenstierna, K. O.
Fox, G. E.
Date Acquired
August 22, 2013
Publication Date
October 1, 1995
Publication Information
Publication: Applied and environmental microbiology
Volume: 61
Issue: 10
ISSN: 0099-2240
Subject Category
Exobiology
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Exobiology

Available Downloads

There are no available downloads for this record.
No Preview Available