Current Issues in Molecular Biology

569 KiB

Open AccessReview

by Diana M. Downs

Curr. Issues Mol. Biol. 2003, 5(1), 17-25; https://doi.org/10.21775/cimb.005.017 - 1 Jan 2003

Cited by 1 | Viewed by 513

The field of bacterial metabolism and physiology is arguably the oldest in microbiology. Much of our understanding of biological processes and molecular paradigms has its roots in early studies of prokaryotic physiology. After a period of declining interest in metabolic studies (prompted by the insurgence of molecular techniques), genomic technologies are revitalizing the study of bacterial metabolism and physiology. These new technologies bring a means to approach metabolic questions with a global perspective. When used in combination with classical and molecular techniques, emerging global technologies will make it feasible to understand the complex integration of metabolic processes that result in an efficient physiology. At the same time, without increased computational capabilities, the massive amounts of data generated by these technologies threaten to overwhelm, rather than facilitate, this work. For genomic technologies to reach their potential for increasing our understanding of bacterial metabolism, microbiologists must become more collaborative and multidisciplinary than at any time in our history. Full article

596 KiB

Open AccessReview

Using Flow Cytometry to Quantify Microbial Heterogeneity

by Hazel M. Davey and Michael K. Winson

Curr. Issues Mol. Biol. 2003, 5(1), 9-15; https://doi.org/10.21775/cimb.005.009 - 1 Jan 2003

Cited by 3 | Viewed by 886

Abstract

Flow cytometry is a powerful technique for the study of single cells, and thus it is of particular utility in the study of heterogeneity in microbial populations. This review seeks to highlight the role of flow cytometric analyses in studies of microbial heterogeneity, drawing wherever possible on recently published research articles. Whilst microbial heterogeneity is well documented in both natural and laboratory environments, the underlying causes are less well understood. Possible sources for the heterogeneity that is observed in microbial systems are discussed, together with the flow cytometric tools that aid its study. The role of flow cytometry in molecular biology is discussed with reference to gene reporter systems, which enable heterogeneity of gene expression to be monitored. With the recent sequencing of a variety of microbial genomes, it is anticipated that flow cytometry will have an increasing role to play in studying the effects of gene expression and mutation on heterogeneity, and in resolving the interactions of genetics and physiology. Full article

657 KiB

Open AccessReview

Direct Extraction of DNA from Soils for Studies in Microbial Ecology

by Mark A. Schneegurt, Sophia Y. Dore and Charles F. Kulpa

Curr. Issues Mol. Biol. 2003, 5(1), 1-8; https://doi.org/10.21775/cimb.005.001 - 1 Jan 2003

Cited by 4 | Viewed by 1997

Abstract

Molecular analyses for the study of soil microbial communities often depend on the extraction of DNA directly from soils. These extractions are by no means trivial, being complicated by humic substances that are inhibitory to PCR and restriction enzymes or being too highly colored for blot hybridization protocols. Many different published protocols exist, but none have been found to be suitable enough to be generally accepted as a standard. Most direct extraction protocols start with relatively harsh cell breakage steps such as bead-beating and freeze-thaw cycles, followed by the addition of detergents and high salt buffers and/or enzymic digestion with lysozyme and proteases. After typical organic extraction and alcohol precipitation, further purification is usually needed to remove inhibitory substances from the extract. The purification steps include size-exclusion chromatography, ion-exchange chromatography, silica gel spin columns, and cesium chloride gradients, among others. A direct DNA extraction protocol is described that has been shown to be effective in a wide variety of soil types. This protocol is experimentally compared to several published protocols. Full article

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Curr. Issues Mol. Biol., Volume 5, Issue 1 (January 2003) – 3 articles

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