In addition, a relationship was demonstrated between the absence of capsule and the incapacity to assign a known serotype to S. suis isolates. We are grateful to K. Kim (Johns Hopkins University School of Medicine) for providing the HBMEC. We wish to thank Sonia Lacouture, Louis Grignon, and Richard Janvier for their technical assistance. This study was supported by the Fond Québécois de la Recherche sur la Nature

BGB324 et les Technologies (FQRNT) and the Natural Sciences and Engineering Research Council of Canada (NSERC). All authors report no conflicts of interest related to their study. “
“In this study, we investigated the β-lactamase-encoding genes responsible for β-lactam resistance phenotypes detected among 56 Gram-negative isolates (Gamma- and Alpha-proteobacteria) recovered PF-2341066 from wastewater, urban streams, and drinking water. The β-lactam resistance mechanisms detected in 36 isolates comprised the presence of class A (blaTEM-1, blaSHV-1, blaSHV-11, blaGES-5), class B (ImiS, L1), class C (blaCMY-2, blaCMY-34, blaCMY-65, blaCMY-89, blaCMY-90, blaACC-5, blaACT-13), and class D (blaOXA-309)β-lactamase-encoding genes, some variants described for the first time here. Notably, the results showed antimicrobial resistance genes related not only to commonly used antibiotics, but also to carbapenems, providing the first

description of a GES-5-producing Enterobacteriaceae. The importance of ubiquitous bacteria thriving in aquatic environments as reservoirs or carriers of clinically relevant resistance determinants was confirmed, and the need to monitor water habitats as potential sources for the emergence and/or spread of antibiotic resistance in the environment was highlighted. “
“Medizinische Fakultät, Klinik für Kinder-Onkologie-Hämatologie und Klinische Immunologie, Heinrich-Heine Universität

Düsseldorf, Düsseldorf, Germany Medizinische Fakultät, Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität München, acetylcholine München, Germany The ubiquitous pathogen Listeria monocytogenes lives either saprophytically in the environment or within cells in a vertebrate host, thus adapting its lifestyle to its ecological niche. Growth experiments at 24 and 37 °C (environmental and host temperature) with ammonium or glutamine as nitrogen sources revealed that ammonium is the preferred nitrogen source of L. monocytogenes. Reduced growth on glutamine is more obvious at 24 °C. Global transcriptional microarray analyses showed that the most striking difference in temperature-dependent transcription was observed for central nitrogen metabolism genes, glnR (glutamine synthetase repressor GlnR), glnA (glutamine synthetase GlnA), amtB (ammonium transporter AmtB), glnK (PII regulatory protein GlnK), and gdh (glutamate dehydrogenase) when cells were grown on glutamine.

In addition, a relationship was demonstrated between the absence of capsule and the incapacity to assign a known serotype to S. suis isolates. We are grateful to K. Kim (Johns Hopkins University School of Medicine) for providing the HBMEC. We wish to thank Sonia Lacouture, Louis Grignon, and Richard Janvier for their technical assistance. This study was supported by the Fond Québécois de la Recherche sur la Nature

selleck chemicals et les Technologies (FQRNT) and the Natural Sciences and Engineering Research Council of Canada (NSERC). All authors report no conflicts of interest related to their study. “
“In this study, we investigated the β-lactamase-encoding genes responsible for β-lactam resistance phenotypes detected among 56 Gram-negative isolates (Gamma- and Alpha-proteobacteria) recovered RAD001 from wastewater, urban streams, and drinking water. The β-lactam resistance mechanisms detected in 36 isolates comprised the presence of class A (blaTEM-1, blaSHV-1, blaSHV-11, blaGES-5), class B (ImiS, L1), class C (blaCMY-2, blaCMY-34, blaCMY-65, blaCMY-89, blaCMY-90, blaACC-5, blaACT-13), and class D (blaOXA-309)β-lactamase-encoding genes, some variants described for the first time here. Notably, the results showed antimicrobial resistance genes related not only to commonly used antibiotics, but also to carbapenems, providing the first

description of a GES-5-producing Enterobacteriaceae. The importance of ubiquitous bacteria thriving in aquatic environments as reservoirs or carriers of clinically relevant resistance determinants was confirmed, and the need to monitor water habitats as potential sources for the emergence and/or spread of antibiotic resistance in the environment was highlighted. “
“Medizinische Fakultät, Klinik für Kinder-Onkologie-Hämatologie und Klinische Immunologie, Heinrich-Heine Universität

Düsseldorf, Düsseldorf, Germany Medizinische Fakultät, Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität München, Enzalutamide nmr München, Germany The ubiquitous pathogen Listeria monocytogenes lives either saprophytically in the environment or within cells in a vertebrate host, thus adapting its lifestyle to its ecological niche. Growth experiments at 24 and 37 °C (environmental and host temperature) with ammonium or glutamine as nitrogen sources revealed that ammonium is the preferred nitrogen source of L. monocytogenes. Reduced growth on glutamine is more obvious at 24 °C. Global transcriptional microarray analyses showed that the most striking difference in temperature-dependent transcription was observed for central nitrogen metabolism genes, glnR (glutamine synthetase repressor GlnR), glnA (glutamine synthetase GlnA), amtB (ammonium transporter AmtB), glnK (PII regulatory protein GlnK), and gdh (glutamate dehydrogenase) when cells were grown on glutamine.

3. This confirms that, under our task’s stimulus conditions, SC inactivation with muscimol did not dramatically alter the temporal patterns of microsaccades commonly observed after the cue. Also note that the saline injection was not associated with the small increase in microsaccade rate observed before cue onset in Fig. 3. This suggests that muscimol in that case did not spread rostrally in the SC, which would be expected to reduce microsaccade rate rather than increase it (Hafed et al.,

2009; Goffart et al., 2012). Finally, when we combined all muscimol injection sessions for the same monkey, we observed a similar pattern of results (Fig. 5A–C): the time course of microsaccades after cue onset was similar to FK228 the pre-inactivation time course, and there was a subtle increase in microsaccade frequency during some epochs. Critically, no evidence for

a reduction of microsaccades was observed in all sessions (even before cue onset with only a single fixation spot on the display), as might be expected from a motor deficit in microsaccade generation if the inactivation had spread to more rostral regions implicated in the motor control of microsaccades (Hafed et al., 2009; Goffart Pexidartinib cell line et al., 2012). Similar analyses of the sessions collected from the second monkey (J) gave similar observations (Fig. 5D–F). Thus, for the stimulus configuration of our task, peripheral SC inactivation did not reduce microsaccade rate, and it did not change the temporal pattern of microsaccades after cue and motion patch onset. Although there was a minimal change in the overall rate of microsaccades, SC inactivation at the peripheral eccentricities associated with our stimuli had a clear effect on the well-known directional biases in microsaccades caused by attentional cueing (Hafed & Clark, 2002; Hafed et al., 2011). We first illustrate this result for the sample Mannose-binding protein-associated serine protease session shown in Fig. 3 by separating movements on the basis of whether they were directed towards the cued location (Fig. 6A, blue rate curves) or towards the foil location

(Fig. 6A, magenta rate curves). Figure 6A also includes ‘raster’ plots of microsaccade onset times, in which the horizontal position of each dot in the raster (x-axis) represents the onset time of a microsaccade, and the vertical position (y-axis) represents trial number. The rasters are color-coded to match the rate curves below them and to identify microsaccades either towards the cued quadrant (blue) or towards the foil quadrant (magenta). For clarity, we did not plot microsaccades directed towards neither the cue nor the foil (the remaining two quadrants of space) in this sample analysis, but we did include these movements in the summary figures described shortly. Before SC inactivation and with the cue placed in the region soon to be affected by muscimol injection (Fig.

multiformis: Nmul; Polaromonas: Pola. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Vibrio cholerae colonizes the human intestine and causes the acute diarrheal disease cholera. Flagellar-mediated

chemotaxis contributes to intestinal colonization as well as infectivity. The virulence-regulatory protein ToxT activates transcription of the genes encoding the major virulence factors cholera toxin and toxin coregulated pilus. ToxT additionally activates transcription of two genes, tcpI and acfB, located within the Vibrio Pathogenicity Island predicted to encode methyl-accepting chemoreceptors. see more We show that disruption of either tcpI or acfB individually does not noticeably affect V. cholerae intestinal colonization within the infant mouse, but disruption of both tcpI and acfB leads to a decrease in intestinal colonization. These results suggest that TcpI and AcfB may have overlapping or redundant chemotactic functions that contribute to V. cholerae intestinal buy SP600125 colonization. Vibrio cholerae causes the acute diarrheal disease cholera in humans. The bacteria are acquired by the ingestion of contaminated water or food, and colonize the intestine. Vibrio cholerae expresses virulence factors

within the intestinal tract that lead to disease symptoms, most notably the cholera toxin (CT), which is responsible for the acute Cyclin-dependent kinase 3 watery diarrhea characteristic of cholera (Holmgren & Svennerholm, 1989). The organisms also express the toxin coregulated

pilus (TCP), a Type IV pilus that is required for intestinal colonization (Taylor et al., 1987). A regulatory cascade commonly referred to as the ToxR regulon coordinately regulates the expression of CT and TCP in response to environmental signals within the host (for a review, see Childers & Klose, 2007). Activation of the ToxR regulon culminates in the expression of the regulatory protein ToxT, which then directly activates transcription of the genes encoding CT and TCP (DiRita et al., 1991). The toxT and tcp genes lie within a cluster of horizontally acquired virulence genes in the Vibrio Pathogenicity Island (VPI) (Karaolis et al., 1998), while the ctx genes are within the lysogenic bacteriophage CTXφ (Waldor & Mekalanos, 1996). Bacterial chemotaxis is accomplished by a number of proteins that constitute a signaling cascade. Methyl-accepting chemoreceptors (MCP) are membrane-spanning proteins that undergo a conformational change upon binding a chemoattractant or a repellant, and this stimulates a signaling cascade that ultimately results in the formation of a phosphorylated chemotaxis protein CheY that interacts with the flagellum and switches the rotation from counterclockwise to clockwise (for a review, see Baker et al., 2006).

8). The infection was newly identified after the initiation of HAART (unmasking IRIS) in eight out of 18 cases

(44.4%). In the remaining 10 cases (55.6%), IRIS was diagnosed after worsening of a previously treated CNS infection (paradoxical IRIS). The median interval from HAART initiation to diagnosis of IRIS was 39 days (IQR 20–90 days). Table 3 shows demographic, clinical and immunological characteristics of patients who developed paradoxical and unmasking IRIS. In order to identify pretreatment variables associated with the risk of developing paradoxical IRIS, these patients were compared with those who did not experience a paradoxical reaction. We found, as the only difference between the two groups, that patients who did not develop IRIS were more likely to have had a previous AIDS-defining condition (51.1% vs. 0% for those developing paradoxical IRIS; P = 0.002). Patients developing IRIS buy NVP-LDE225 had a more rapid immunological recovery than patients who did

not develop IRIS, as evidenced by a greater increase in CD4 count after Cyclopamine 3 months of antiretroviral therapy (ART) (170 vs. 62 cells/μL, respectively; P < 0.025). At this time-point, the decrease in viral load was also greater among patients with paradoxical IRIS, but differences did not reach statistical significance (–2.6 vs. −1.8 log10 for those with paradoxical IRIS; P = 0.10). Patients who began HAART within CHIR-99021 supplier 2 weeks after the diagnosis of a CNS infection were not at higher risk of developing paradoxical IRIS (50% vs. 65.8% for those who began HAART more than 2 weeks after diagnosis; P = 0.32). Figure 3 shows the cumulative probabilities of survival and the median survival time categorized by the development and type of IRIS. We did not find significant differences in survival between patients who developed paradoxical IRIS and those without IRIS. Eight (44.4%) of the 18 patients with IRIS received therapy

with steroids for a variable period depending on the response to therapy and other individual patient characteristics. None of the 10 patients who were not treated with steroids died, while three of the eight who received steroids died. In those three cases, mortality was directly attributed to IRIS. These three patients had PML. In our study, we observed a progressive decline in the incidence of CNS opportunistic infections during the first decade of the 21st Century. The overall rate of CNS infections decreased significantly from 8.3 cases per 1000 HIV-infected patients in the year 2000 to 1.4 in 2010. Since HAART became available, many studies have reported a decrease in the incidence of most opportunistic conditions related to HIV infection, including neurological infections [1-6, 20-22]. For example, a study performed in France by Abgrall et al. in 2001 showed a reduction of 34% in the risk of cerebral toxoplasmosis after the introduction of protease inhibitors [5].

, 2001). C/EBP β, especially LAP1 and LAP2, can be phosphorylated at several sites by many different protein kinases, such as mitogen-activated PD-1 antibody protein kinases, protein kinase A, protein kinase C, glycogen synthase kinase 3, and calcium/calmodulin-dependent protein kinases, with different effects on its transcriptional activity, depending on the phosphorylation site (Mahoney et al., 1992; Wegner et al., 1992; Trautwein et al., 1993, 1994; Piwien-Pilipuk et al., 2001, 2002). In particular, whereas phosphorylation

of rat C/EBP β by protein kinase A, protein kinase C or glycogen synthase kinase 3 on Ser240, which is located in the DNA-binding domain, has been reported to attenuate DNA binding and induce nuclear export, Ser105 phosphorylation of LAP isoforms is a key determinant of its transactivation capacity (Trautwein et al., 1993, 1994; Buck et al., 1999; Piwien-Pilipuk et al., 2001, see more 2002). We therefore evaluated C/EBP β phosphorylation on Ser105 as a marker of transcriptional activity for this transcription factor. By using an antibody that specifically recognizes C/EBP β phosphorylated on Ser105, we observed that LAP1 is phosphorylated on Ser105 only in the nuclear compartment, implying

its transcriptional activation. From our co-immunoprecipitation experiments, we determined that LAP1 is essentially present in CGNs in its sumoylated form, both in the cytoplasm and in the nucleus, Anacetrapib and that the phosphorylated form is only nuclear and is only detected when neurons are kept in pro-survival conditions. The SUMOs serve as modifiers, exerting their effect by becoming conjugated to target proteins and stabilizing them (reviewed

by Lieberman, 2004). Sumoylation provides a rapid and efficient way to modulate the subcellular localization, activity and stability of a wide variety of protein substrates (Dorval & Fraser, 2007). C/EBPs, including C/EBP β, are well-known targets of SUMOs, which control their transcriptional activity by releasing, in rats, the inhibitory action of a conserved inhibitory domain that is a target for lysine sumoylation (Kim et al., 2002). Concerning C/EBP β isoforms, both LAP1 and LAP2 are potential targets of SUMO-2/3, but only LAP1 has been demonstrated to be conjugated to SUMO-2/3, as confirmed by our present results in CGNs. C/EBP β sumoylation has been shown to regulate its transcriptional activity, without influencing its subcellular localization (Eaton & Sealy, 2003). When CGNs were shifted to K5 medium to induce apoptosis, we observed a decrease in the LAP1 level and an increase in the LIP level in the nuclear compartment, and a decrease in the LAP2 level in the cytosolic fraction. Concomitantly, p-(Ser105)-LAP1 disappeared from the nuclear fraction.

This

suggests that HIV-1 may be constrained in its ability to become both highly resistant and highly fit, and that reduced viral fitness is an important factor contributing to persistent partial suppression of viral replication during long-term virological failure. Our results, exploring the predictive value of RC in the largest published cohort of treatment-experienced patients undergoing treatment selleck screening library interruption, did not support these hypotheses. As reviewed by Martinez-Picado and Martinez [30], the association of RC and plasma HIV-1 RNA levels in patients with viraemia has been found to be either weak [31] or limited to small pilot cohorts [18]. There are limited published data on the predictive value of RC in treatment-experienced

patients in the HAART era [22]. Our results for the predictive value of PSS are in accordance with those of Lawrence et al. [32] and Katzenstein et al. [33] in demonstrating that PSS predicts early virological response to salvage HAART. Potential limitations of our study include the large diversity in treatment regimens that patients in our cohort received, PLX3397 solubility dmso given the fact that OPTIMA was a strategy trial rather than a specific combination regimen trial. We also did not account for the possibility of secondary failures resulting in alterations in the salvage regimens, but these are less likely to occur within the first 12 weeks. We limited our analysis to this early phase for this reason, and also because of the attrition in the number of

patients with samples available at the later time-points, which would limit our ability to assess the predictive value of both PSS and RC values at baseline. In summary, in this large cohort of ARV treatment-experienced patients undergoing different salvage treatment strategies, our results confirm that RC increases Edoxaban after treatment interruption and baseline PSS predicts changes in viraemia both during treatment interruption and early in salvage therapy. iPSS did not have a better predictive value than dPSS. The latter, easier measurement should be evaluated in predicting responses to the newer classes of ARVs. We further demonstrated that baseline RC does not predict changes in CD4 cell count during either treatment interruption or salvage therapy and does not provided added information to PSS. We thank all the subjects, investigators and staff who participated in the OPTIMA trial. This study was funded by the Department of Veterans Affairs Cooperative Studies Program and, in part, by a Department of Veterans Affairs grant to MH, and by Monogram BioSciences Inc., who provided support for performance of the PhenoSense™ and replication capacity assays.

This

suggests that HIV-1 may be constrained in its ability to become both highly resistant and highly fit, and that reduced viral fitness is an important factor contributing to persistent partial suppression of viral replication during long-term virological failure. Our results, exploring the predictive value of RC in the largest published cohort of treatment-experienced patients undergoing treatment Selleckchem CYC202 interruption, did not support these hypotheses. As reviewed by Martinez-Picado and Martinez [30], the association of RC and plasma HIV-1 RNA levels in patients with viraemia has been found to be either weak [31] or limited to small pilot cohorts [18]. There are limited published data on the predictive value of RC in treatment-experienced

patients in the HAART era [22]. Our results for the predictive value of PSS are in accordance with those of Lawrence et al. [32] and Katzenstein et al. [33] in demonstrating that PSS predicts early virological response to salvage HAART. Potential limitations of our study include the large diversity in treatment regimens that patients in our cohort received, Staurosporine ic50 given the fact that OPTIMA was a strategy trial rather than a specific combination regimen trial. We also did not account for the possibility of secondary failures resulting in alterations in the salvage regimens, but these are less likely to occur within the first 12 weeks. We limited our analysis to this early phase for this reason, and also because of the attrition in the number of

patients with samples available at the later time-points, which would limit our ability to assess the predictive value of both PSS and RC values at baseline. In summary, in this large cohort of ARV treatment-experienced patients undergoing different salvage treatment strategies, our results confirm that RC increases (-)-p-Bromotetramisole Oxalate after treatment interruption and baseline PSS predicts changes in viraemia both during treatment interruption and early in salvage therapy. iPSS did not have a better predictive value than dPSS. The latter, easier measurement should be evaluated in predicting responses to the newer classes of ARVs. We further demonstrated that baseline RC does not predict changes in CD4 cell count during either treatment interruption or salvage therapy and does not provided added information to PSS. We thank all the subjects, investigators and staff who participated in the OPTIMA trial. This study was funded by the Department of Veterans Affairs Cooperative Studies Program and, in part, by a Department of Veterans Affairs grant to MH, and by Monogram BioSciences Inc., who provided support for performance of the PhenoSense™ and replication capacity assays.

The cells for SEM observation were critical-point dried and applied to a silicon wafer slide. The cells were then examined using a JSM-6360 scanning electron microscope (JEOL) (Qu et al., 2008). The cells (grown at 42 °C for 144 h) for TEM observation were embedded in the Epon 812 embedding kit and cut into ultrathin sections. The sections were double-stained with uranyl acetate and lead nitrate and then examined using a JEM-2000EX TEM (JEOL). The lipopolysaccharides

prepared from MV501 (pYJ), MV501 (pYJ-1), MV501 (pYJ-2) and MV501 (pUC18) cells were analyzed by SDS-PAGE, followed by silver staining (Fig. 2). The lipopolysaccharides from MV501 (pYJ), MV501 (pYJ-1) and MV501 (pYJ-2) cells showed a ladder-like banding pattern buy Veliparib characteristic of O side-chain material. The results suggested that Rv1302 and MSMEG_4947 have the same function as E. coli WecA and both Rv1302 and MSMEG_4947 utilize C55-P and UDP-GlcNAc as substrates to HDAC inhibitors list initiate the synthesis of the O7 polysaccharide that is covalently linked to the lipid A-core oligosaccharide of E. coli O7:K1 strain VW187 (Valvano & Crosa, 1989). The MSMEG_4947 in conditional replication plasmid pYJ-4 was disrupted by inserting a kanR cassette.

A two-step homologous recombination procedure (Li et al., 2006) was used to achieve the allelic replacement of the MSMEG_4947 gene by MSMEG_4947∷kanR. MSMEG_4947 (406 amino acids) shares 79% identity with Rv1302 (404 amino acids); therefore, the rescue plasmid pYJ-6 carrying Rv1302 was constructed for complementation studies. The MSMEG_4947 knockout mutant was confirmed by a Southern blot Dichloromethane dehalogenase using MSMEG_4947 as a probe (Fig. 3). The growth of five MSMEG_4947 knockout mutants (nos 1–5) was investigated at both 30 and 42 °C. All five MSMEG_4947 knockout mutants had similar growth patterns and the growth curve of no. 2 mutant is shown in Fig. 4a. The results clearly showed that the MSMEG_4947 knockout mutant grew only at 30 °C and not at 42 °C. The rescue plasmid pYJ-6 was unable to replicate at 42 °C and, therefore, no more Rv1302 protein was generated. In contrast, wild-type mc2155 containing

pCG76 grew at both 30 and 42 °C, confirming that MSMEG_4947 was essential for the growth of M. smegmatis. To investigate whether a decrease in WecA has effects on the morphology of the MSMEG_4947 knockout mutant, a certain amount of cells was acquired by performing a temperature shift experiment. The MSMEG_4947 knockout mutant (no. 2) was grown at 30 °C for 24 h to produce Rv1302 protein, and then grown at 42 °C. A600 nm was measured at 24-h intervals (Fig. 4b) and the cells were harvested for observation of the morphological phenotype (Fig. 5). MSMEG_4947 knockout cells grown at 30 °C for 72 (Fig. 5a) and 144 h (Fig. 5c) had a smooth cell surface and exhibited the normal rod-like shape seen in the wild-type mc2155 cells (Qu et al., 2008).

The cells for SEM observation were critical-point dried and applied to a silicon wafer slide. The cells were then examined using a JSM-6360 scanning electron microscope (JEOL) (Qu et al., 2008). The cells (grown at 42 °C for 144 h) for TEM observation were embedded in the Epon 812 embedding kit and cut into ultrathin sections. The sections were double-stained with uranyl acetate and lead nitrate and then examined using a JEM-2000EX TEM (JEOL). The lipopolysaccharides

prepared from MV501 (pYJ), MV501 (pYJ-1), MV501 (pYJ-2) and MV501 (pUC18) cells were analyzed by SDS-PAGE, followed by silver staining (Fig. 2). The lipopolysaccharides from MV501 (pYJ), MV501 (pYJ-1) and MV501 (pYJ-2) cells showed a ladder-like banding pattern see more characteristic of O side-chain material. The results suggested that Rv1302 and MSMEG_4947 have the same function as E. coli WecA and both Rv1302 and MSMEG_4947 utilize C55-P and UDP-GlcNAc as substrates to ICG-001 ic50 initiate the synthesis of the O7 polysaccharide that is covalently linked to the lipid A-core oligosaccharide of E. coli O7:K1 strain VW187 (Valvano & Crosa, 1989). The MSMEG_4947 in conditional replication plasmid pYJ-4 was disrupted by inserting a kanR cassette.

A two-step homologous recombination procedure (Li et al., 2006) was used to achieve the allelic replacement of the MSMEG_4947 gene by MSMEG_4947∷kanR. MSMEG_4947 (406 amino acids) shares 79% identity with Rv1302 (404 amino acids); therefore, the rescue plasmid pYJ-6 carrying Rv1302 was constructed for complementation studies. The MSMEG_4947 knockout mutant was confirmed by a Southern blot Florfenicol using MSMEG_4947 as a probe (Fig. 3). The growth of five MSMEG_4947 knockout mutants (nos 1–5) was investigated at both 30 and 42 °C. All five MSMEG_4947 knockout mutants had similar growth patterns and the growth curve of no. 2 mutant is shown in Fig. 4a. The results clearly showed that the MSMEG_4947 knockout mutant grew only at 30 °C and not at 42 °C. The rescue plasmid pYJ-6 was unable to replicate at 42 °C and, therefore, no more Rv1302 protein was generated. In contrast, wild-type mc2155 containing

pCG76 grew at both 30 and 42 °C, confirming that MSMEG_4947 was essential for the growth of M. smegmatis. To investigate whether a decrease in WecA has effects on the morphology of the MSMEG_4947 knockout mutant, a certain amount of cells was acquired by performing a temperature shift experiment. The MSMEG_4947 knockout mutant (no. 2) was grown at 30 °C for 24 h to produce Rv1302 protein, and then grown at 42 °C. A600 nm was measured at 24-h intervals (Fig. 4b) and the cells were harvested for observation of the morphological phenotype (Fig. 5). MSMEG_4947 knockout cells grown at 30 °C for 72 (Fig. 5a) and 144 h (Fig. 5c) had a smooth cell surface and exhibited the normal rod-like shape seen in the wild-type mc2155 cells (Qu et al., 2008).