Use of a Plasmid DNA Probe To Monitor Populations of Bacillus pumilus Inoculant Strains in Hay

Autor:	Carol A. Hendrick, Brenda K. Smiley, Tomes Nancy J, Terrence H. Shelley
Rok vydání:	1991
Předmět:	Bacillaceae Ecology biology Bacillus pumilus fungi Genetics and Molecular Biology Molecular cloning biology.organism_classification Applied Microbiology and Biotechnology Restriction fragment Microbiology Plasmid Hay biology.protein Molecular probe Food Science Biotechnology Southern blot
Zdroj:	Applied and Environmental Microbiology. 57:686-693
ISSN:	1098-5336 0099-2240
DOI:	10.1128/aem.57.3.686-693.1991
Popis:	We are evaluating naturally occurring isolates of Bacillus pumilus for use as microbial hay preservatives. Seven isolates of B. pumilus from hay contained a 42-kb cryptic plasmid (pMGD296). We wished to determine whether pMGD296 could be used as a molecular marker to follow populations of these isolates in hay over time. Southern blots and colony blots of 69 isolates of B. pumilus and other Bacillus spp. were probed with 32 P-labeled pMGD296. Twenty-nine probe-positive isolates were identified; of these, 28 contained a plasmid with a restriction profile identical to that of pMGD296. One isolate from untreated hay contained a 40-kb plasmid (pMGD150) that was homologous to pMGD296 but had a different restriction fragment pattern. Regions of homology between the two plasmids were identified by Southern blotting, and a 1.9-kb Hin dIII- Pst I fragment of pMGD296 lacking strong homology to pMGD150 was cloned in pUC18. The cloned fragment hybridized only with isolates containing pMGD296 and was used to estimate populations of these isolates in treated and untreated hay.
Databáze:	OpenAIRE
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