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Title: ANALYSIS OF THE EFFECTS OF TEMPERATURE SHIFTS ON LEPTOSPIRAL GENE EXPRESSION AS ASSESSED BY WHOLE GENOME MICROARRAY

Author
item LO, MIRANDA - MONASH UNIVERSITY
item BULACH, D - MONASH UNIVERSITY
item HAAKE, D - UCLA
item MATSUNAGA, J - UCLA
item Paustian, Michael
item Zuerner, Richard
item ADLER, B - MONASH UNIVERSITY

Submitted to: International Leptospirosis Society Meeting Abstracts and Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 6/18/2005
Publication Date: 11/13/2005
Citation: Lo, M., Bulach, D.M., Haake, D.A., Matsunaga, J., Paustian, M., Zuerner, R.L., Adler, B. 2005. Analysis of the effects of temperature shifts on leptospiral gene expression as assessed by whole genome microarray [abstract]. International Leptospirosis Society Meeting Abstracts and Proceedings. p.143.

Interpretive Summary:

Technical Abstract: Background: Presently, little is known about gene regulation or expression profiles of Leptospira grown under different environmental conditions. Since the genomes of three different strains have been completely annotated, whole genome microarray technology can now be applied to examine transcriptional changes in Leptspira grown under a range of conditions. Of particular interest in this study are temperature-responsive genes since leptospires are routinely grown in vitro at 30°C, but can clearly grow in the environment and in vivo at temperatures of 37°C and higher. Methods: Oligonucleotide arrays, representing 3605 open reading frames (ORFs) from L. interrogans sv Lai plus an additional 45 ORFs unique to sv Copenhageni, were used to assess transcriptional profiles of L. interrogans sv Lai grown at 20°C, 30°C, 37°C and 39°C. Results: Poor annotation of the L. interrogans sv Lai genome has hampered genetic analysis. Therefore, the genome had to be re-annotated prior to design of the whole genome array. The original annotation of the Lai genome had 4768 ORFs which we reduced to 3666 ORFs in our revised annotation. The revised annotation was subsequently used in the design of oligonucleotides for the whole genome array. Oligonucleotides of 70 bases in length were selected using the program OligoArraySelector. Pilot studies indicated that a range of genes was differentially expressed by at least 2-fold across the range of temperatures at which L. interrogans sv Lai was grown. Conclusions: Global changes in L. interrogans sv Lai gene expression were observed as a result of growth under different temperatures. Keywords: Genome annotation, gene expression, microarray, temperature regulation