5 and 97.3% retention of viability after incubation in serum, respectively, compared to 9% viability of serovar Patoc. However, after incubation with
heat-inactivated serum (HIS) the viability of L. biflexa was greater than 95%, consistent with the killing effect of serum being due to complement activity. Accordingly, serovar Copenhageni was used in subsequent microarray experiments, since microarray slides were constructed based on the combined complete genome sequences S3I-201 nmr of serovars Lai and Copenhageni available in the database . Global transcriptomic changes of pathogenic Leptospira after serum exposure Low-passage L. interrogans serovar Copenhageni was incubated with 50% guinea pig serum at 37°C for 30 min to simulate in vivo conditions encountered upon entry into the host. Comparisons were made with leptospires shifted to 37°C in EMJH medium to exclude the effect of temperature shift, which has previously been reported [10, 11]. Overall, 168 genes (4.5% of the genome) were considered to be differentially expressed
at a statistically significant level upon serum exposure, i.e. at least 1.5-fold up- or down-regulated with an adjusted P value of less than 0.01 as determined by moderated t test. Of these, 55 genes (32.7%) were up-regulated and 113 genes (67.3%) were down-regulated (Table 1). Genes of known or predicted function accounted for 54.5% (30 of 55 genes) and 45.1% (51 of 113 genes) of up- and down-regulated genes, respectively. Table 1 Number of leptospiral genes differentially KPT-8602 chemical structure expressed in response to serum compared to EMJH medium Genes No. of genes Up-regulated (%a) Down-regulated TSA HDAC manufacturer (%a) Total (%b) Known or predicted function 30 (54.5) 51 (45.1) 81 (48.2) Unknown or poorly characterized function 25 (45.5) 62 (54.9) 87 (51.8) Total 55 113 168 a percentage of genes per total number of genes in up-regulated or down-regulated group b percentage of genes per total number of differentially expressed genes Differentially expressed genes were classified into functional categories based on clusters of orthologous groups (COGs). The majority of differentially expressed genes Adenosine were of poorly characterized
or unknown function (45.5 and 54.9% of up- and down-regulated genes, respectively) (Figure 1A). In general, of the genes which were serum-inducible, those predicted to be involved in metabolism were overrepresented, followed by the cellular processes and signaling group (Figure 1A). However, down-regulated genes of known or predicted function were similarly distributed in three broad COG categories. Among genes of known or predicted function, the highest proportion of up-regulated genes (10.9%) were those involved in cell wall and membrane biogenesis (COG category M), whereas the largest group of down-regulated genes (11.5%) belonged to COG category J (translation) (Figure 1B). Figure 1 Percentage of up- and down-regulated genes of L.