POTENTIAL RISKS OF GENE AMPLIFICATION BY PCR AS DETERMINED BY 16S RDNA ANALYSIS 
OF A MIXED-CULTURE OF STRICT BAROPHILIC BACTERIA

Liesack, W; Weyland, H; Stackebrandt, E

doi:10.1007/BF02539153

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POTENTIAL RISKS OF GENE AMPLIFICATION BY PCR AS DETERMINED BY 16S RDNA ANALYSIS OF A MIXED-CULTURE OF STRICT BAROPHILIC BACTERIA

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Citation

Liesack, W., Weyland, H., & Stackebrandt, E. (1991). POTENTIAL RISKS OF GENE AMPLIFICATION BY PCR AS DETERMINED BY 16S RDNA ANALYSIS OF A MIXED-CULTURE OF STRICT BAROPHILIC BACTERIA. MICROBIAL ECOLOGY, 21(3), 191-198. doi:10.1007/BF02539153.

Cite as: https://hdl.handle.net/21.11116/0000-000D-6BAD-9

Abstract

The 16S rDNA genes of an apparently pure culture of a psychrophilic and
strict barophilic bacterium (WHB 46) were studied by PCR-mediated
amplification and cloning into phage M13 mp 18. Sequence analysis of
five individual clones revealed the presence of two different 16S rDNA
types. The homology value of 90% indicates that culture WHB 46 is
actually composed of two closely related species (WHB 46-1 and 46-2).
Both strains are members of the gamma-subdivision of proteobacteria.
Analysis of a sixth clone (WHB 46-1/2) leads to the conclusion that it
represents a 16S rDNA hybrid molecule assembled during the PCR reaction.
This hypothesis was confirmed by secondary structure analysis of the
chimeric rDNA. The appearance of such hybrid molecules point to a
potential risk in studies on the diversity of bacterial populations by
analysis of rDNA pattern via PCR-mediated amplification because they
suggest the existence of organisms that do not actually exist in the
sample investigated.