One hundred and forty soil samples, collected from 30-cm soil depth in Nampong District, Khon Kaen Province, Thailand, were used for phage isolation using the basic enrichment method PF-562271 mouse (Kutter & Sulakvelidze, 2005). Five grams of soil were inoculated into 20 mL of brain–heart infusion broth (Oxoid, Basingstoke, UK), mixed and incubated at 37 °C for 16–18 h.

Five milliliters of the culture were centrifuged at 4000 g, 4 °C for 30 min and the supernatant filtered through 0.22-μm filters and used as phage lysate or stored at 4 °C until use. The spot test method was used to screen for the presence of lytic phage activity (Chopin et al., 1976). Approximately 1 mL of mid-log phase B. pseudomallei P37 (1 × 109 CFU mL−1) was flooded onto a plate containing nutrient agar with 3.6 mM CaCl2, the excess removed and allowed to dry open in a laminar flow biosafety cabinet. Twenty microliters of phage lysate from each soil sample were then dropped

onto the plate and incubated at 37 °C overnight and the clear zone formation was observed. Each clear and isolated plaque was cored out by a sterile Pasteur pipette into nutrient broth, shaken for 1 h and centrifuged at 2500 g, at 4 °C for 20 min. Supernatants were filtered through 0.22-μm filter membranes and purified by the PI3K inhibitor soft agar method (Sambrook & Russell, 2001). The purification steps for each phage were repeated three times to ensure the homogeneity of the phage stock and finally phage titers were calculated as PFU mL−1. A Phosphoglycerate kinase mid-log phase culture of B. pseudomallei P37 (1 × 109 CFU mL−1) in 100 mL nutrient broth (Oxoid) containing 3.6 mM CaCl2 was mixed with the purified phage suspension at a multiplicity of infection (MOI) of 0.1 and incubated at 37 °C for 3–5 h. After bacterial lysis was observed, the solution was centrifuged and the supernatant containing phage particles was filtered through

0.22-μm filter membranes and used as the phage suspension. One hundred microliters of each B. pseudomallei isolate’s overnight culture were spread on the surface of nutrient agar plates and 20 μL of each phage suspension (∼108–109 PFU mL−1) was spotted and incubated at 37 °C for 18–24 h. The results were recorded as negative if there were no plaques and positive if clear plaques were observed. The host range of selected phages was further evaluated with species closely related to Burkholderia (Table 1) by the agar overlay method (Sambrook & Russell, 2001). The negative staining method was performed to visualize phage morphology using transmission electron microscopy (Jamalludeen et al., 2007). Ten microliters of phage suspension (>108 particles mL−1) were used for staining with 10 μL of 2% uranyl acetate for 10 min. Photographs were taken under a transmission electron microscope (JEM-2100, JEOL LAB6, Japan). Size was determined from the average of three independent measurements.

One hundred and forty soil samples, collected from 30-cm soil depth in Nampong District, Khon Kaen Province, Thailand, were used for phage isolation using the basic enrichment method Talazoparib solubility dmso (Kutter & Sulakvelidze, 2005). Five grams of soil were inoculated into 20 mL of brain–heart infusion broth (Oxoid, Basingstoke, UK), mixed and incubated at 37 °C for 16–18 h.

Five milliliters of the culture were centrifuged at 4000 g, 4 °C for 30 min and the supernatant filtered through 0.22-μm filters and used as phage lysate or stored at 4 °C until use. The spot test method was used to screen for the presence of lytic phage activity (Chopin et al., 1976). Approximately 1 mL of mid-log phase B. pseudomallei P37 (1 × 109 CFU mL−1) was flooded onto a plate containing nutrient agar with 3.6 mM CaCl2, the excess removed and allowed to dry open in a laminar flow biosafety cabinet. Twenty microliters of phage lysate from each soil sample were then dropped

onto the plate and incubated at 37 °C overnight and the clear zone formation was observed. Each clear and isolated plaque was cored out by a sterile Pasteur pipette into nutrient broth, shaken for 1 h and centrifuged at 2500 g, at 4 °C for 20 min. Supernatants were filtered through 0.22-μm filter membranes and purified by the http://www.selleckchem.com/products/3-methyladenine.html soft agar method (Sambrook & Russell, 2001). The purification steps for each phage were repeated three times to ensure the homogeneity of the phage stock and finally phage titers were calculated as PFU mL−1. A Dapagliflozin mid-log phase culture of B. pseudomallei P37 (1 × 109 CFU mL−1) in 100 mL nutrient broth (Oxoid) containing 3.6 mM CaCl2 was mixed with the purified phage suspension at a multiplicity of infection (MOI) of 0.1 and incubated at 37 °C for 3–5 h. After bacterial lysis was observed, the solution was centrifuged and the supernatant containing phage particles was filtered through

0.22-μm filter membranes and used as the phage suspension. One hundred microliters of each B. pseudomallei isolate’s overnight culture were spread on the surface of nutrient agar plates and 20 μL of each phage suspension (∼108–109 PFU mL−1) was spotted and incubated at 37 °C for 18–24 h. The results were recorded as negative if there were no plaques and positive if clear plaques were observed. The host range of selected phages was further evaluated with species closely related to Burkholderia (Table 1) by the agar overlay method (Sambrook & Russell, 2001). The negative staining method was performed to visualize phage morphology using transmission electron microscopy (Jamalludeen et al., 2007). Ten microliters of phage suspension (>108 particles mL−1) were used for staining with 10 μL of 2% uranyl acetate for 10 min. Photographs were taken under a transmission electron microscope (JEM-2100, JEOL LAB6, Japan). Size was determined from the average of three independent measurements.

glutamicum

is generally recognized as a nonhazardous organism, and thus safe to handle. Furthermore, its central metabolism has been extremely well investigated and there are well-established molecular biology tools for manipulation, so C. glutamicum is a particularly suitable model organism for mycolic acid-containing actinomycetes. The complete genome sequence of C. glutamicum ATCC 13032 was determined, and predicted to contain 3002 ORFs, with the function of 2489 of these identified by homologies to known proteins (Kalinowski et al., 2003). A blastp search has revealed AP24534 price that M. tuberculosis, Mycobacterium bovis and C. glutamicum have intact thyA gene, and a gene with strong similarity to thyX. Amino acid sequence alignments revealed a fully conserved ThyX motif (RHRX7S) common to this protein. The ThyX of C. glutamicum exhibited 63% identity in amino acid sequence to that of M. tuberculosis. However, the reason why both of these genes are maintained in these organisms is not yet understood. In the present study, we developed a C. glutamicum mutant lacking

thyX. This demonstrated that thyX is not essential for active growth and that its absence makes the organism more sensitive to WR99210, an active triazine inhibitor of DHFR. We also carried out a long-term starvation study that revealed that the survival of a thyX mutant of C. glutamicum was greatly impaired during stationary growth phase. The bacterial strains are listed in Table 1. Escherichia coli and C. glutamicum selleckchem strains were cultured at 37 °C in Luria–Bertani (LB) medium and at 30 °C in nutrient broth. Minimal media for both E. coli and C. glutamicum were M9 and MCGC (Minimum Corynebacterium glutamicum Citrate) (Von der Osten et al., 1989), with glucose added to a final concentration of 1% w/v. Ampicillin (100 μg mL−1), kanamycin (25 μg mL−1)

and WR99210 (20 μM) were added to the media when required. The predicted genes were identified by 72% and 63% sequence similarity at amino acid level to M. tuberculosis ThyA and ThyX, respectively. PCR was used to amplify the coding sequence of the thyA and thyX genes from C. glutamicum ATCC 13032. The DNA fragment clonidine corresponding to the thyA gene was amplified using primers THYA1 and THYA2, and the thyX DNA fragment of C. glutamicum was amplified using oligonucleotides THYX1 and THYX2. The PCR fragments were cloned into the plasmid pUC18 and sequenced to verify the accuracy of the clones. An E. coliχ2913 strain lacking thyA was used as the host for transformation (Dower et al., 1988): transformation was performed by electroporation of pUC18 containing thyA and pUC18 containing thyX. Escherichia coliχ2913 transformants, carrying thyA or thyX from C. glutamicum, were streaked on M9 minimal agar in the absence of thymidine, and retained for further experimentation.

These results suggest that nonassociative plasticity modifies neural networks in such a way that it affects local competitive Dabrafenib purchase interactions among mixture components. We used a computational model to evaluate the most likely targets for modification. Hebbian modification of synapses from inhibitory

local interneurons to projection neurons most reliably produced the observed shift in response to the mixture. These results are consistent with a model in which the antennal lobe acts to filter olfactory information according to its relevance for performing a particular task. “
“Neuronal networks in the spinal cord termed central pattern generators (CPGs) are responsible for the generation of rhythmic movements, such as walking. The axon guidance molecule EphA4 has been suggested to play a role in the configuration of spinal CPG networks in mammals. In EphA4 knockout (EphA4-KO) mice, the normal alternating walking pattern is replaced by a rabbit-like hopping gait, which Omipalisib manufacturer can be reproduced

when locomotor-like activity is induced in the isolated spinal cord. This hopping phenotype has been explained by an abnormal midline crossing of ipsilateral axons. Here, we investigated the nature of this overcrossing in heterozygous EphA4 (EphA4lacZ/+) mice that showed normal alternating gait and homozygous EphA4 (EphA4lacZ/lacZ) mice with hopping gait. Localized lesions showed that the hopping phenotype is maintained by fibers crossing in the ventral commissure. Using transgenic mouse lines in which glutamatergic, GABAergic

and glycinergic neurons are intrinsically labeled, we showed a significant increase Venetoclax clinical trial in the number of crossing excitatory β-galactosidase-positive neurons and a decrease in the number of inhibitory neurons crossing the midline in EphA4lacZ/lacZ mice compared with EphA4lacZ/+ mice. These results show that the hopping phenotype is the result of a change in the balance between excitatory and inhibitory signals across the midline and that EphA4-positive neurons play an essential role in the mammalian CPG. “
“Visual expertise in discriminating fine differences among a group of similar objects can be obtained through extensive long-term training. Here we investigated the neural bases of this superior capability. The inferotemporal cortex, located at the final stage along the ventral visual pathway, was a candidate site in monkeys because cells there respond to various complex features of objects. To identify the changes that underlie the development of visual expertise in fine discrimination, we created a set of parametrically designed object stimuli and compared the stimulus selectivity of inferotemporal cells between two different training histories.

These results suggest that nonassociative plasticity modifies neural networks in such a way that it affects local competitive PLX3397 in vivo interactions among mixture components. We used a computational model to evaluate the most likely targets for modification. Hebbian modification of synapses from inhibitory

local interneurons to projection neurons most reliably produced the observed shift in response to the mixture. These results are consistent with a model in which the antennal lobe acts to filter olfactory information according to its relevance for performing a particular task. “
“Neuronal networks in the spinal cord termed central pattern generators (CPGs) are responsible for the generation of rhythmic movements, such as walking. The axon guidance molecule EphA4 has been suggested to play a role in the configuration of spinal CPG networks in mammals. In EphA4 knockout (EphA4-KO) mice, the normal alternating walking pattern is replaced by a rabbit-like hopping gait, which Carfilzomib purchase can be reproduced

when locomotor-like activity is induced in the isolated spinal cord. This hopping phenotype has been explained by an abnormal midline crossing of ipsilateral axons. Here, we investigated the nature of this overcrossing in heterozygous EphA4 (EphA4lacZ/+) mice that showed normal alternating gait and homozygous EphA4 (EphA4lacZ/lacZ) mice with hopping gait. Localized lesions showed that the hopping phenotype is maintained by fibers crossing in the ventral commissure. Using transgenic mouse lines in which glutamatergic, GABAergic

and glycinergic neurons are intrinsically labeled, we showed a significant increase Farnesyltransferase in the number of crossing excitatory β-galactosidase-positive neurons and a decrease in the number of inhibitory neurons crossing the midline in EphA4lacZ/lacZ mice compared with EphA4lacZ/+ mice. These results show that the hopping phenotype is the result of a change in the balance between excitatory and inhibitory signals across the midline and that EphA4-positive neurons play an essential role in the mammalian CPG. “
“Visual expertise in discriminating fine differences among a group of similar objects can be obtained through extensive long-term training. Here we investigated the neural bases of this superior capability. The inferotemporal cortex, located at the final stage along the ventral visual pathway, was a candidate site in monkeys because cells there respond to various complex features of objects. To identify the changes that underlie the development of visual expertise in fine discrimination, we created a set of parametrically designed object stimuli and compared the stimulus selectivity of inferotemporal cells between two different training histories.

, 1999). BIME-1 and BIME-2 correspond to SMAG TT and HH dimers. However, HH dimers are about 10 times more abundant than TT dimers. In contrast, BIME-1 (74 repeats) are three times more abundant than BIME-2 (24 repeats).

Moreover, both BIME-1 and BIME-2 are invariably comprised of elements from different subfamilies (Bachellier et al., 1999; see also http://www.pasteur.fr/recherche/unites/pmtg/repet/index.html). The predominance learn more of TT over HH dimers, and the composite nature of dimers, is also a distinctive feature of the abundant REP families found in Pseudomonas putida (Aranda-Olmedo et al., 2002) and P. syringae (Feil et al., 2005). It has been hypothesized that REPs are mobilized by a click here transposase of the IS200/IS605 family, and the corresponding genes have been shown to be flanked by REPs in many species (Nunvar et al., 2010). Four genes encoding this transposase were identified in K279a DNA (ORFs 1101,

1152, 2816 and 4509), but only ORFs 1101 and 2816 are flanked by SMAGs. We believe that REPs are an ancient component of the genomes of Proteobacteria, which have been actively mobilized by transposition only early in their history. According to this view, REPs disappeared in time from most species, their dissemination being plausibly detrimental to the cell, and have been maintained only in species in which they could no longer transpose. This hypothesis is supported by the observation that SMAG sequences were found in none of the 41 species-specific GEIs, plausibly acquired by lateral gene transfer, which account for >10% of the K279a chromosome (Rocco et al., 2009). REPs are similarly restricted to core genome regions in P. syringae (Tobes & Pareja, 2005). In contrast to what was observed for REPs in other species (Tobes & Pareja, 2006), SMAGs are not targeted by mobile DNA. However, it is worth noting that a K279a GEI encoding type 1 pili (Rocco et al., 2009) is flanked by SMAG-2 dimers. BCKDHB About 1/7 of the ORFs of the K279a strain are flanked by SMAGs in a distance range that makes the presence of promoter or terminator

sequences unlikely. It is plausible that most of these elements are transcribed into mRNA, and that their folding into RNA hairpins may influence the level of expression of flanking genes. The number of genes potentially controlled at the post-transcriptional level by SMAGs may be higher than estimated, because many repeats are inserted either upstream (17 elements) or downstream (150 elements) or within (44 elements) known or putative operons. We analyzed genes transcribed in the same direction intermingled with SMAG sequences, and found that the repeats influence the segmental mRNA stability. Both monomers and dimers function as stabilizers of upstream transcripts, and work with comparable efficiency when embedded in the same RNA context (Fig. 5).

, 1999). BIME-1 and BIME-2 correspond to SMAG TT and HH dimers. However, HH dimers are about 10 times more abundant than TT dimers. In contrast, BIME-1 (74 repeats) are three times more abundant than BIME-2 (24 repeats).

Moreover, both BIME-1 and BIME-2 are invariably comprised of elements from different subfamilies (Bachellier et al., 1999; see also http://www.pasteur.fr/recherche/unites/pmtg/repet/index.html). The predominance this website of TT over HH dimers, and the composite nature of dimers, is also a distinctive feature of the abundant REP families found in Pseudomonas putida (Aranda-Olmedo et al., 2002) and P. syringae (Feil et al., 2005). It has been hypothesized that REPs are mobilized by a Talazoparib solubility dmso transposase of the IS200/IS605 family, and the corresponding genes have been shown to be flanked by REPs in many species (Nunvar et al., 2010). Four genes encoding this transposase were identified in K279a DNA (ORFs 1101,

1152, 2816 and 4509), but only ORFs 1101 and 2816 are flanked by SMAGs. We believe that REPs are an ancient component of the genomes of Proteobacteria, which have been actively mobilized by transposition only early in their history. According to this view, REPs disappeared in time from most species, their dissemination being plausibly detrimental to the cell, and have been maintained only in species in which they could no longer transpose. This hypothesis is supported by the observation that SMAG sequences were found in none of the 41 species-specific GEIs, plausibly acquired by lateral gene transfer, which account for >10% of the K279a chromosome (Rocco et al., 2009). REPs are similarly restricted to core genome regions in P. syringae (Tobes & Pareja, 2005). In contrast to what was observed for REPs in other species (Tobes & Pareja, 2006), SMAGs are not targeted by mobile DNA. However, it is worth noting that a K279a GEI encoding type 1 pili (Rocco et al., 2009) is flanked by SMAG-2 dimers. Mirabegron About 1/7 of the ORFs of the K279a strain are flanked by SMAGs in a distance range that makes the presence of promoter or terminator

sequences unlikely. It is plausible that most of these elements are transcribed into mRNA, and that their folding into RNA hairpins may influence the level of expression of flanking genes. The number of genes potentially controlled at the post-transcriptional level by SMAGs may be higher than estimated, because many repeats are inserted either upstream (17 elements) or downstream (150 elements) or within (44 elements) known or putative operons. We analyzed genes transcribed in the same direction intermingled with SMAG sequences, and found that the repeats influence the segmental mRNA stability. Both monomers and dimers function as stabilizers of upstream transcripts, and work with comparable efficiency when embedded in the same RNA context (Fig. 5).

To validate these observations, we experimentally converted the mutL gene between the wild-type and 6bpΔmutL in S. typhimurium and inspected the bacterial mutability status. When 6bpΔmutL was converted

to mutL, the originally highly mutable Salmonella strains regained genetic stability; when mutL was converted to 6bpΔmutL, the mutability was elevated 100-fold. Interestingly, ERK inhibitor libraries mutL cells were found to grow out of 6bpΔmutL cells; the new mutL cells eventually replaced the original 6bpΔmutL population. As conversion between mutL and 6bpΔmutL may occur readily during DNA replication, it may represent a previously unrecognized mechanism to modulate bacterial mutability at the population level, allowing bacteria to respond rapidly to changing environments while minimizing the risks associated with persistent hypermutability. Previously, we observed the peculiar phenomenon of genome diversification, i.e. mutants of Salmonella MS-275 solubility dmso typhimurium LT7 stored at room temperature kept changing the physical structure of the genome (Liu et al., 2003). We have since concentrated on identifying the genetic basis responsible for this phenomenon, with a focus on mismatch repair (MMR) genes including mutL, mutS and mutH.

Our initial work showed that all screened S. typhimurium LT7 mutants had intact mutS and mutH genes, but a deletion was found in mutL; this genotype was designated 6bpΔmutL (Gong et al., 2007). MutL has been suggested to function as a master coordinator or molecular matchmaker in the MMR system. It has a weak ATPase selleck kinase inhibitor function, binds to DNA, interacts directly with MutS, MutH and UvrD, and is required for initiation as well as subsequent steps in MMR processes (Ban & Yang, 1998; Hall et al., 1998; Ban et al., 1999; Spampinato & Modrich, 2000). The deleted sequence that we identified in mutL is one of three tandem 6-bp

repeats, GCTGGC GCTGGC GCTGGC. Similar repeats were reported in Escherichia coli, although they were presented as G CTGGCG CTGGCG CTGGCG (Shaver & Sniegowski, 2003). The sequence (G)CTGGC GCTGGC GCTGGC C in Salmonella and (G) CTGGCG CTGGCG CTGGCG in E. coli both code for the amino acid sequence LALALA, which lies in a region of MutL that forms the lid of the ATP-binding pocket (Ban & Yang, 1998; Ban et al., 1999; Yang, 2000; Yang et al., 2000). In our case, the deletion of one of the 6-bp repeats will lead to one of the three LA sets missing in the ATP lid structure of MutL and may thus impair ATPase activity. As the repeat structure would facilitate deletion or duplication via slipped-strand mispairing (Streisinger et al., 1966; Levinson & Gutman, 1987), one can imagine that MMR may cease functioning when the 6-bp repeats of mutL decrease or increase one or more copies and resume functioning when the copy number again becomes three, both by ‘errors’ during replication.

The increased generation of ROS at the tissue level induces a wide range of biological Protein Tyrosine Kinase inhibitor activity such as lipid peroxidation, protein denaturation,

inactivation of enzymes and decomposition of cellular DNA.[70] In this way, ROS may cause cellular and tissue damage. These unwanted effects of ROS may cause impairment of ova or sperm function. Bacterial endotoxin-induced increase in ROS production may also cause caspase-mediated apoptosis.[69] This apoptosis-inducing effect of ROS may result in endometrial or tubal epithelial damage, and impairment in fertilization and sperm motility.[62, 63] We now know that innate immunity plays an important role in the initiation of immune response in the pelvic environment. A number of

widely accepted mechanisms involved in the development or pathogenesis of endometriosis are summarized and shown in Figure 3. The production of pro-inflammatory cytokines and growth of endometriosis in the pelvic TAM Receptor inhibitor environment can be regulated by the innate immune system. We proposed for the first time a new concept ‘bacterial contamination hypothesis’ in endometriosis and involvement of LPS/TLR4 cascade in the growth regulation of endometriosis. Our results suggest that a substantial amount of endotoxin in peritoneal fluid due to reflux of menstrual blood is involved in pelvic inflammation and may promote TLR4-mediated growth of endometriosis. Targeting bacterial endotoxin, TLR4 or NF-κB could be useful as a therapeutic strategy to suppress pelvic inflammation and growth of endometriosis with consequent improvement in the quality of life and fertility rate of women who suffer from this enigmatic disease. Our ongoing study to find evidence of a subclinical infection within the vaginal cavity of women with endometriosis may hold new PRKD3 therapeutic potential in addition to conventional estrogen-suppressing agent. A complete understanding of the mechanisms of the innate immunity and TLR system will be helpful for the future development of innovative

therapies for the manipulation of endometriosis and other reproductive diseases. We thank Miss Kazumi Hayashida and Miss Kyoko Ishida, Department of Obstetrics and Gynecology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan, for their excellent technical assistance. This work was supported by Grants-in-Aid for Scientific Research (no. 16591671 and 18591837) from the Ministry of Education, Sports, Culture, Science and Technology of Japan (to K. N. K.). None declared. “
“Shakuyaku-kanzo-to, a Kampo medicine composed equally of shakuyaku and kanzo, is an antispasmodic drug that can inhibit contraction of uterine smooth muscles in pregnant women and rats. We aimed to test the inhibitory effects of water- and lipid-soluble extracts of shakuyaku-kanzo-to, shakuyaku, and kanzo in order to identify the fraction responsible for inhibiting uterine smooth muscle contraction in pregnancy.

, 2007). Notably, the phenotypic effects of the absence of DnaE2 appear more clearly in P. putida mutant lacking DNA Pol I, indicating that DnaE2 may complement in part some functions of Pol I. It is known that Pol I participates in the gap-filling

reaction in the NER pathway. Unpublished results in our laboratory show that the Pol I mutant of P. putida is less sensitive to UV irradiation than P. putida lacking the NER system, which indicates that some other DNA polymerase could perform DNA repair synthesis in NER when Pol I is missing. Additional deletion of the dnaE2 gene in the Pol I-deficient P. putida reduces the UV tolerance of bacteria and increases the mutation frequency, Staurosporine supplier whereas the viability of UV-irradiated DnaE2-deficient bacteria is not reduced when Pol I is present. These results imply that DnaE2 may partially complement the absence of Pol I in a DNA damage repair pathway such as NER. Additionally, because the mutation frequency

is lower in UV-irradiated DnaE2-proficient cells than in those lacking Bleomycin DnaE2, TLS carried out by this DNA polymerase might be accurate. In contrast to the results obtained with P. putida DnaE2, Sanders et al. (2006) have demonstrated that UV-induced mutagenesis in P. aeruginosa is dependent on Pol I and DnaE2, i.e., the mutation frequency was decreased when measured in UV-irradiated P. aeruginosa transposon library mutants either carrying insertions in Pol I or DnaE2 genes. These genetic data suggest that P. aeruginosa DnaE2, different from its P. putida homologue, is mutagenic. Thus, DnaE2s from P. putida and P. aeruginosa would provide a good model to study the molecular mechanisms influencing the fidelity of DnaE2 homologues. According to its sequence similarity, P. putida ImuB and its homologues form a branch in the UmuC superfamily of proteins that is distinct from E. coli-like DinB proteins (Pol IV) (Galhardo et al., 2005). However, the absence of conserved residues forming a

catalytic center of Y-family polymerases in ImuB raises a question of whether ImuB has a DNA polymerase activity at all (Koorits et al., 2007). So far, the exact role of ImuB in Pseudomonas species has remained unclear. Deletion of the dnaE2 gene from ImuB-deficient P. putida did not increase the mutation frequency (Koorits et al., 2007), thereby Tenofovir molecular weight suggesting that ImuB might be needed for DnaE2 activity. Genetic data obtained in other organisms such as C. crescentus indicate that ImuB possibly cooperates with DnaE2 in DNA damage-inducible mutagenesis, as no phenotypic effect of DnaE2 was demonstrated in this organism in the absence of ImuB (Galhardo et al., 2005). The question is whether ImuB could assist only DnaE2. The possibility that ImuB may cooperate not only with DnaE2, but could also influence the activity of other DNA polymerases is supported by the finding that deletion of only the imuB or the dnaE2 gene from P.