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A biphasic approach to the determination of the phenotypic and genotypic diversity of some anaerobic, cellulolytic, thermophilic, rod-shaped bacteria.

A biphasic approach, involving a numerical phenetic and a phylogenetic study, was used to determine diversity among some anaerobic, cellulolytic, thermophilic, rod-shaped bacteria. Ninety two characters were determined for 51 strains in the numerical taxonomy study, and partial 16S rDNA sequences from 16 isolates were compared. Both the phenetic and phylogenetic data indicate diversity within this group of organisms, and reveal the lack of similarity between sporogenous and asporogenous isolates. The results of the phylogenetic study demonstrate the lack of relationship of the majority of the strains studied to previously studied thermophilic bacteria. In general, good correlation exists between the two data sets, but discrepancies arise when strains with a high level of similarity are examined. The need for caution in the interpretation of data obtained from such a biphasic approach is discussed.

Anti-Bacterial Agents↗
PubMed1996

Cloning vectors for lactococci based on a plasmid encoding resistance to cadmium.

An 8.8-kb plasmid (pND302) was identified in Lactococcus lacti spp lactis M71 which encodes cadmium resistance (CdR). Most of the commercial lactococcal strains tested were sensitive to cadmium. Therefore, CdR should provide a useful selectable marker for constructing cloning vectors in lactococci. pND302 was mapped with a number of restriction enzymes and found to contain a unique EcoRI site suitable for cloning. Two E. coli/L lactis shuttle cloning vectors, pND304 and pND624, were constructed by subcloning of the E. coli plasmids pBR322 and pGEM-7Zf(+) containing a 1.6-kb gene encoding nisin resistance (NisR) of lactococcal origin into the EcoRI site of pND302, separately. The E. coli DNA component of pND624 was removed and the resulting plasmid, pND625, consisted of only lactococcal DNA, expressing NisR and CdR, with two synthetic polylinkers that contain multiple restriction sites for versatile cloning. Both pND302 and pND625 can be transformed by electroporation into L. lactis LMO230 at 10(3)/micrograms DNA and maintained stably in LMO230. The results indicated that pND302 and pND625 are potential food-grade cloning vectors for lactococci.

Base Sequence↗