Cloning, sequencing, and characterization of genomic subtracted sequences from Listeria monocytogenes

Article Abstract:

Genomic subtracted sequences from Listeria monocytogenes, a food-borne pathogen, have been characterized. Individual sequences of a genomic subtracted, PCR-amplified mixed-sequence probe (GS probe) were cloned and sequenced. The GS probe differentiated restriction fragment length polymorphism (RFLP) pattern for L. monocytogenes, and did not hybridize with members of other genera. Some sequences were already in the GenBank database and some had no significant level of homology. Some putative identities of other sequences were inferred from homology data. Subtractive hybridization is convenient for isolating multiple unique DNA sequences that may be in two nucleic acid pools.

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Isolation of laccase gene-specific sequences from white rot and brown rot fungi by PCR

Article Abstract:

The genetic basis of fungal degradation of lignin by the enzyme laccase from white rot and brown rot fungi was investigated by polymerase chain reaction. Analysis of PCR-amplified products showed a DNA band of approximately 200 base pairs in six genera. A significant homology in amino acid sequences was seen in the PCR products of the white rot fungi Ganoderma lucidum, Phlebia brevispora and Trametes versicolor, but not in Lentinula edodes and Lentinus tigrinus. The nucleotide sequence similarity for the three genera of white rot fungi was 65 to 74% while that for the brown rot fungi Gloeophyllum trabeum was 60 to 71%.

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Combined subtraction hybridization and polymerase chain reaction amplification procedure for isolation of strain-specific Rhizobium DNA sequences

Article Abstract:

A procedure for the isolation of strain-specific Rhizobium leguminosarum bv. trifoli DNA sequences was reported. The probe strain DNA was hybridized with subtracter DNA derived from a group of related strains. The subtraction system utilized a combination of four strategies: separation of probe strain and subtracter DNA by streptavidin-phenol-chloroform extraction; desalting of probe strain DNA and removal of remaining biotinylated DNA by NENSORB chromatography; destruction of subtracter DNA by UDG; and amplification by polymerase chain reaction with a probe strain-specific primer.

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