Rapid method for direct extraction of mRNA from seeded soils

Journal Article · Thu Feb 28 23:00:00 EST 1991 · Applied and Environmental Microbiology; (United States)

OSTI ID:5273573

Tsai, Yuli; Park, M J; Olson, B H ^[1]

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1. Univ. of California, Irvine (USA)

Subsurface soils from an old gas manufacturing site in southern California were used for RNA extraction. The contaminated soils contained high concentrations of polynuclear aromatic hydrocarbons (6.5 {mu}g/g of soil) and toxic metals. The total CFU of heterotrophic bacteria in both samples were enumerated with plate count agar (PCA). The isolated mercury- or methyl mercury-resistant strains were able to grow on PCA amended with 25 {mu}g of Hg{sup 2+} (as HgCl{sub 2}) per ml or 3 {mu}g of CH{sub 3}-Hg{sup +} (as CH{sub 3}(Hg)OH) per ml, respectively, and the naphthalene degraders were isolated from minimal medium saturated with naphthalene vapor. The method described extracts up to 17 {mu}g of total RNA per g (wet weight) of soil containing 8.0 {times} 10{sup 8} cells of Pseudomonas aeruginosa PU21, and mRNA has been detected in 160-ng total RNA fractions. This method has been used for the detection of mRNA transcribed from specific biodegradative genes, including the nah and mer operons, in contaminated soils. This extraction method can be completed within a few hours and has tremendous potential for ecological studies of in situ gene expression among soil microbiotas.

• OSTI ID: 5273573
• Journal Information: Applied and Environmental Microbiology; (United States), Journal Name: Applied and Environmental Microbiology; (United States) Vol. 57:3; ISSN 0099-2240; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

54 ENVIRONMENTAL SCIENCES
540220* -- Environment
Terrestrial-- Chemicals Monitoring & Transport-- (1990-)
550200 -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AROMATICS
BACTERIA
BIOASSAY
CFU
CONDENSED AROMATICS
CONTAMINATION
ELEMENTS
GENE OPERONS
HYDROCARBONS
MERCURY
MERCURY COMPOUNDS
MESSENGER-RNA
METALS
METHYLMERCURY
MICROORGANISMS
MOLECULAR BIOLOGY
NAPHTHALENE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC MERCURY COMPOUNDS
POLYCYCLIC AROMATIC HYDROCARBONS
PSEUDOMONAS
RNA
SOILS