Bacterial artificial chromosome fingerprint arrays for the differentiation of transcriptomic differences in mycobacteria

Journal of Microbiological Methods
Alice H LiRichard W Stokes

Abstract

Although microarray technology has become more widespread as a discovery tool for bacterial pathogenesis, it remains a method available only to laboratories with access to expensive equipment and costly analysis software. Mycobacterium tuberculosis, the causative agent for tuberculosis (TB), afflicts one-third of the global population, and kills between 2 and 3 million people per year. While the majority of cases of TB occur in developing areas of the world, facilities in these regions may not be able to support microarray analysis. Additionally, a major limitation of microarrays is that only genes on the array are being assayed. With acquired virulence and drug resistance in microbes, a method less dependent on a predetermined list of gene targets is advantageous. We present a method of expression analysis based on bacterial artificial chromosomes (BACs) that can be applied with standard laboratory equipment and free analysis software. This technique, bacterial artificial chromosome fingerprint arrays (BACFA), was developed and utilised to identify expression differences between intracellular strains of M. tuberculosis, one virulent (H37Rv) and one attenuated (H37Ra). Southern blots of restriction-enzyme digested BAC fragments...Continue Reading

References

Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·J E Graham, J E Clark-Curtiss
Oct 18, 2002·Nucleic Acids Research·Adel M TalaatStephen Albert Johnston
Oct 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sarah A StanleyJeffery S Cox
Jun 23, 2004·Tuberculosis·D G Niranjala MuttucumaruTanya Parish

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