Chapter 13 : Plasmids

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This chapter reviews what is known concerning plasmids of the genus . It includes methods for plasmid isolation, analysis of plasmid distribution, and identification of plasmid-encoded genes. It also discusses the background for use of mycobacterial plasmids as vectors. There are several obstacles to isolation and characterization of mycobacterial plasmids. The first is the slow growth rate of most mycobacteria. Second, all mycobacterial cells are difficult to lyse, making efficient isolation of intact circular plasmid DNA difficult. Third, mycobacterial cell walls contain complex lipids and polysaccharides that can contaminate DNA preparations, rendering them unsuitable for molecular biologic techniques. The demonstration of related plasmids in different , , and strains suggests that mycobacterial plasmids are capable of horizontal transfer. The permeability barrier of mycobacteria that results in resistance to antibiotics, intracellular killing by macrophages, and environmental stresses is likely to limit the ability of DNA to traverse the cell wall and membrane. Introduction of kanamycin resistance or other antibiotic resistance genes into various mycobacterial plasmids and the use of electroporation to introduce the plasmids into other strains may allow analysis of plasmid functions.

Citation: Falkinham J, Crawford J. 1994. Plasmids, p 185-198. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch13
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