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Measuring the absolute abundance of the microbiome by adding yeast containing 16S rRNA gene from a hyperthermophile

 Ju Yeong Kim  ;  Myung-hee Yi  ;  Myungjun Kim  ;  Seogwon Lee  ;  Hye Su Moon  ;  Dongeun Yong  ;  Tai-Soon Yong 
 MICROBIOLOGYOPEN, Vol.10(4) : e1220, 2021-08 
Journal Title
Issue Date
16S rRNA gene ; absolute abundance ; hyperthermophile ; microbiome ; Thermus aquaticus
High-throughput sequencing (HTS) of 16S rRNA gene amplicons provides compo sitional information regarding the microbial community, but not the absolute abun dance of the bacteria. We aimed to develop a standardized method for quantifying
the absolute abundance of bacteria in microbiome studies. To demonstrate the utility
of our approach, we quantified the number of bacteria from the compositional data of
the fecal and cecal microbiomes. The 16S rRNA gene of a hyperthermophile, Thermus
aquaticus, was cloned into Pichia pastoris (yeast) genome, and an equivalent amount of
the yeast was added to the stool and cecal samples of mice before DNA extraction.
16S rRNA gene library construction and HTS were performed after DNA extraction.
The absolute abundances of bacteria were calculated using T. aquaticus reads. The
average relative abundances of T. aquaticus in the five stool and five cecal samples
were 0.95% and 0.33%, respectively, indicating that the number of bacteria in a cecum
sample is 2.9 times higher than that in a stool sample. The method proposed for quan tifying the absolute abundance of the bacterial population in this study is expected
to overcome the limitation of showing only compositional data in most microbiome
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1. College of Medicine (의과대학) > Dept. of Tropica Medicine (열대의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Kim, Ju Yeong(김주영) ORCID logo https://orcid.org/0000-0003-2456-6298
Yong, Dong Eun(용동은) ORCID logo https://orcid.org/0000-0002-1225-8477
Yong, Tai Soon(용태순) ORCID logo https://orcid.org/0000-0002-3445-0769
Yi, Myung Hee(이명희) ORCID logo https://orcid.org/0000-0001-9537-5726
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