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Chapter 15 : Homologous Recombination, DNA Repair, and Mycobacterial Genes

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Homologous Recombination, DNA Repair, and Mycobacterial Genes, Page 1 of 2

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Abstract:

This chapter discusses the mechanisms involved in homologous recombination and DNA repair, what is known about these systems in mycobacteria, and recent information on the unusual structure of the gene in the pathogenic mycobacteria. The RecA protein of induces the expression of several genes in response to DNA damage, resulting in an increase in cell survival. The RecA protein is essential for homologous recombination in . Homologous recombination provides a tool for creating and characterizing specific mutations. "Gene knockout" techniques, for example, involve the replacement of a wild-type gene with a disrupted, nonfunctional mutated gene. Homologous recombination has been reported in the fast-growing mycobacterium . When DNA encoding the intein of was used to probe genomic DNA of a range of mycobacteria, no hybridization was obtained. The presence of independently acquired inteins in the genes of the two major mycobacterial pathogens raised the possibility that they are a common feature of mycobacterial genes. Recent studies of the genes of mycobacteria have indicated that the major human pathogens and have an unusual structure, suggesting that these organisms may have evolved novel mechanisms for dealing with DNA damage and effecting genetic recombination.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Figure 1

Multiple roles of the RecA protein following exposure to DNA damage.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Figure 2

Role of RecA protein in regulation of the SOS response.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Image of Figure 3
Figure 3

Role of RecA protein in homologous recombination, ds, double stranded; ss, single stranded.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Image of Figure 4
Figure 4

Homologous recombination as a laboratory tool. Homologous recombination can be used to delete genes by incorporating a disrupted gene, in which case the sequence used to disrupt the gene is usually an antibiotic resistance gene or other selectable marker (a), or to introduce a mutation by double crossover events (b).

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Figure 5

(a) Comparison of the M. tuberculosis and E. coli RecA proteins, (b) Production of the mature M. tuberculosis protein by posttranslational splicing.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Image of Figure 6
Figure 6

Known examples of inteins (protein introns). The conserved hexapeptide motif at the intein-C-extein junction is shown in expanded form, with identity to the Saccharomyces cenevisiae VMA1 sequence highlighted. The Candida tropicalis VMA1 gene is described in Gu et al., 1993.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Figure 7

Schematic comparison of the M. leprae and M. tuberculosis recA gene structures. The insertion sites of the inteins within the genes are shown along with the nucleotide sequences flanking the insertion sites. The sizes, sequences, and insertion sites of the two inteins differ, indicating the independent origins of the two genetic elements.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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Figure 8

Inteins are confined to the genes of and the complex. Primers to conserved sequences were used in PCR reactions on genomic DNA from the various mycobacteria, (a) The size of the PCR product reveals the presence or absence of an intein. (b) Only BCG, and show long products, (c) All clinical isolates of had an intein. The primers used in panels b and c differed and hence gave products of different sizes in the two experiments.

Citation: Colston M, Davis E. 1994. Homologous Recombination, DNA Repair, and Mycobacterial Genes, p 217-226. In Bloom B (ed), Tuberculosis. ASM Press, Washington, DC. doi: 10.1128/9781555818357.ch15
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References

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