Carbon-13 (13C) labeling of Bacillus subtilis vegetative cells and spores: suitability for DNA stable isotope probing (DNA-SIP) of spores in soils.
To test the suitability of DNA stable isotope probing (DNA-SIP) for characterizing bacterial spore populations in soils, the properties of Bacillus subtilis cells and spores intensely labeled with [(13)C]glucose were characterized. Spore germination, vegetative growth rates, and sporulation efficiency were indistinguishable on glucose versus [(13)C]glucose, as were spore wet heat and UV resistance. Unlabeled and (13)C-labeled spores contained 1.0989 and 74.336 at.% (13)C, and exhibited wet densities of 1.356 and 1.365 g/ml, respectively. Chromosomal DNAs containing (12)C versus (13)C were readily separated by their different buoyant densities in cesium chloride/ethidium bromide gradients.
Nicholson
Wayne L
WL
Department of Microbiology and Cell Science, University of Florida, Space Life Sciences Laboratory, Kennedy Space Center, FL 32899, USA. WLN@ufl.edu
Fedenko
Jeffrey
J
Schuerger
Andrew C
AC
D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D002247:Carbon Isotopes / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics; D012988:Soil Microbiology; D013171:Spores, Bacterial / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D013194:Staining and Labeling
D002247:Carbon Isotopes; D004269:DNA, Bacterial; D002244:Carbon