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Chapter 96 : Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria

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Abstract:

Many toxic metals appear to be able to enter cells by import systems for essential elements or molecules, and although chromosomally encoded homeostasis systems for essential metals are widespread, in bacteria many toxic metal resistance genes are found on mobile genetic elements such as plasmids and transposons. Highthroughput methods in DNA sequencing, metagenomics, and the postgenomic technologies are going to revolutionize our understanding of toxic metal-bacterial cell interactions and the diversity of toxic metal(loid) resistances. With this in mind, this chapter talks about the options and potential workflows open to researchers wishing to use molecular techniques for the study of these resistance genes. Most of the techniques discussed in the chapter are equally applicable to the study of other nonmetal resistance phenotypes of environmental bacteria, because the techniques are generic. The chapter, however, focuses on and discusses the merits of molecular biology techniques that can be used to investigate bacterial resistance mechanisms. The outline of the experimental procedure and primary purpose of this technique is to culture toxic-metal-resistant bacteria, so that the genetic and biochemical basis for their resistance mechanisms can be further investigated. Although we are now entering a new phase in molecular research into toxic metal resistances, the techniques described in this chapter still have a valuable place in describing new metal-resistant bacteria.

Citation: Hobman J. 2007. Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria, p 1166-1182. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch96

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Mobile Genetic Elements
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Gene Expression and Regulation
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Agarose Gel Electrophoresis
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FIGURE 2

Diagrammatic representation of some metal resistance operons from bacteria: ( ), ( ), ( ), ( ), ( ), ( ), and ( ).

Citation: Hobman J. 2007. Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria, p 1166-1182. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch96
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Image of FIGURE 3
FIGURE 3

(a) Isolation and molecular techniques for the analysis of culturable toxic-metal-resistant bacteria. (b) Isolation and molecular techniques for the analysis of toxic metal resistance genes isolated directly from environmental samples. Modified from references and with permission.

Citation: Hobman J. 2007. Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria, p 1166-1182. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch96
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Tables

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Citation: Hobman J. 2007. Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria, p 1166-1182. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch96
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TABLE 1

PCR primers used to amplify specific resistance genes from toxic-metal-resistant bacteria

Citation: Hobman J. 2007. Molecular Techniques for the Study of Toxic Metal Resistance Mechanisms in Bacteria, p 1166-1182. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch96

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