Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 1/26/2006
Publication Date: 1/26/2006
Citation: Paustian, M., Bannantine, J.P. 2006. An oligonucleotide microarray designed for functional genomic analyses of Mycobacterium avium subspecies paratuberculosis [abstract]. Poster No. 2. Interpretive Summary: The bacterium Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) causes Johne’s Disease, an economically important intestinal infection of ruminants such as cattle and sheep. The work described here details the construction of a DNA microarray consisting of short pieces of DNA spotted onto a specially coated glass slide. The DNA sequences used on the microarray represent all of the genes identified from the genome sequence of M. paratuberculosis as well as related sequences from the genome of Mycobacterium avium subspecies avium. The microarrays were produced with funds leveraged from the Johne’s Disease Integrated Program and are therefore available to other researchers. Other scientists should benefit from the results of this work. They will be able to utilize the DNA microarray to perform a wide range of experiments designed to study the ability of M. paratuberculosis to cause disease.
Technical Abstract: The genome sequence of Mycobacterium avium subspecies paratuberculosis (Map) K10 was used to design a microarray of 70mer oligonucleotides. All of the predicted protein coding sequences and intergenic regions from Map K10 as well as novel predicted protein coding sequences from Mycobacterium avium subspecies avium (Maa) 104 are represented on the microarray. Additionally, the microarray contains sheared Map K10 genomic DNA and oligonucleotides representing Map K10 16s rRNA and six Arabidopsis thaliana genes as controls. The microarrays are printed on poly-l-lysine coated glass slides and every oligonucleotide is spotted in triplicate, resulting in 16,350 total spots per slide. These microarrays have thus far been successfully utilized for both comparative genomic hybridizations to identify genetic differences between Map isolates and for transcriptional analyses designed to monitor gene expression in response to environmental conditions. For example, hybridization of fluorescently labeled genomic DNA from bovine and ovine Map isolates identified seven large sequence polymorphisms and numerous smaller sequence changes. The microarrays are available for researchers through the JDIP Genomics, Antibodies and Proteomics Core.