The ratio of male to female participants differed between the two groups. To ensure that the reported group effects were not

driven by gender differences, we also performed the above analyses without the female participants. For all but one test, the pattern of significant results was the same. In the case of the peripheral VEP P1, the amplitude difference between ASD and TD groups approached significance (t30 = 1.87, P = 0.072). As this trended in the predicted direction, Fluorouracil and the other tests replicated the main analyses, we interpret the data based on the main analyses. The current study examined visual processing of central and peripheral inputs in ASD children and adolescents. We hypothesized that their peripheral processing might be altered, as they often exhibit peculiarities in eye-fixation and eye-movement behavior, which probably influence the development of peripheral cortical visual representations. Bleomycin Under this hypothesis, processing of centrally fixated inputs should be largely unaffected, and indeed we found indistinguishable responses between TD and ASD groups for central stimulation for all stimulus types employed. This is not fully consistent with prior reports, as processing differences for central inputs have been reported (Boeschoten et al., 2007; Neumann et al., 2011). Notably, eye position is usually

not tightly controlled, as it was here. Thus, differences in cortical representation for different areas of space, or more variability in eye position in one group over the other, could partially account for these differences. In contrast to responses to centrally presented stimuli, we did uncover marked differences in visual responses to stimuli presented to peripheral

portions of the retina, a finding replicated Phosphoprotein phosphatase across all three stimulus conditions. These peripheral differences reached significance in the timeframe of the P1, indicative of changes in early extrastriate visual areas during relatively early sensory–perceptual processing timeframes (Di Russo et al., 2002; Foxe & Simpson, 2002). The electrophysiological response in the P1 timeframe is generated by multiple visual cortical areas including V1, V2, V3 and V4 (Di Russo et al., 2002). On the other hand, the simple cortical magnification model introduced earlier is entirely based on measurements in V1. Nonetheless, work has shown a general maintenance of spatial mapping patterns across progressively higher levels of the cortical hierarchy, such that one would expect initially reorganized spatial maps to be maintained to at least some degree in later retinotopically mapped regions (Motter, 2009; Harvey & Dumoulin, 2011), although as receptive field sizes progressively increase along the hierarchy, an entirely strict one-to-one maintenance of initial mapping would seem unlikely.

(2005), M. silvestris prefers acetate over methane as a growth substrate, possibly due

to the requirement of reducing equivalents for the initial oxidation of methane to methanol, and because acetate concentrations can be quite high in Sphagnum peat bogs where this strain was isolated. As a result it appears that facultative methanotrophic Methylocella strains have an effective regulatory network to control MMO expression. Conversely, the facultative Methylocystis strains H2s and SB2 were found to constitutively express pMMO regardless if these strains were grown on methane or acetate (Belova et al., 2011; Yoon et al., 2011). Expression of pmoA, a key functional gene of the pMMO however, was significantly greater when Methylocystis strain SB2 was grown on methane than on acetate (Yoon et al., 2011). As described above, these strains show weaker growth on acetate. It may be that these strains BMS-907351 concentration use acetate as

a secondary carbon or reducing source that enables the continued expression of MMO in the absence of methane such that these strains can readily utilize methane when it becomes more available (Dunfield, 2007; Belova et al., 2011). These strains, however, were also isolated from bogs where acetate concentrations can be expected to be high (Duddleston et al., 2002), thus, the ability to control MMO expression may have other origins. It is interesting to note that sMMO expression in Methylococcus capsulatus Bath is repressed at high copper concentrations, while pMMO is constitutively expressed but its expression increases with increasing copper concentration (Choi Alectinib et al., 2003). The finding that sMMO expression by M. silvestris is repressed in the presence of acetate while pMMO expression is constitutive and positively regulated by the carbon source in Methylocystis strain SB2 suggests that the regulatory pathway of sMMO/pMMO expression used by facultative

and obligate methanotrophs have some similarities. It may be that Methylocella species were originally facultative methylotrophs, later generating the ability to utilize methane as a growth substrate through lateral gene transfer of the genes for the sMMO, and subsequently developed the ability to control MMO expression with respect to carbon source. This is intriguing as Methylocella species are the only known methanotrophs lacking pMMO. By extension, M. aurea may also have been methylotrophic, but through find more lateral gene transfer developed the ability to express pMMO. To the best of our knowledge, however, it should be stressed that it has not yet been reported whether M. aurea expresses pMMO when grown on acetate. Although the origin of facultative methanotrophy from methylotrophs in these strains is speculative, it is interesting to note that a facultative methylotroph, Methylobacterium extorquens AM1, when engineered to express the ammonia monooxygenase (AMO) of Paracoccus denitrificans, was able to grow on methane as the sole carbon source (Crossman et al., 1997).

, 2008). Besides being implicated in dimer formation, the conserved cysteine residue is of interest because a mutational analysis of certain motif residues in E. coli Ygf Z implicated C228 as a determinant of plumbagin

sensitivity (Lin et al., 2010). To gain further insight into the function of the Ygf Z motif, this study analysed the criticality of each of its conserved residues to growth and to MiaB activity. Only C228 was found to be indispensable. Complementation studies were carried out using the ΔygfZ strain described previously (Waller et al., 2010). This strain was transformed with pBAD24 selleck containing the wild-type E. coli ygfZ gene (EcYgfZ∷pBAD24; Waller et al., 2010) or mutants thereof, in which one of the conserved residues in the Ygf Z motif had been replaced by alanine by site-directed mutagenesis (Cormack, 2008). Cells were grown at 37 °C in Antibiotic Medium 3 (Difco), LB medium with or without 30 μM plumbagin, or M9 minimal medium plus 2 g L−1 glycerol as indicated. Media were solidified with 15 g L−1 agar; ampicillin and kanamycin were used at 50 and 50 μg mL−1, respectively. Gene expression was induced with 0.2 g L−1 l-arabinose. Growth kinetics were followed in a Bioscreen-C Automated Growth Curve Analysis System (Growth Curves Obeticholic Acid ic50 USA, MA) using the following parameters: continuous shaking; reading every 30 min; culture volume, 200 μL. As inoculum, overnight cultures in LB plus ampicillin

and kanamycin (washed three times with M9 Clostridium perfringens alpha toxin medium plus glycerol before dilution) were diluted to give a final OD600 nm of 0.005. Bioscreen experiments used triplicate cultures of three independent strains. Bulk nucleic acids were isolated from stationary phase cells cultured in Antibiotic Medium 3 and enriched for tRNA (Bailly et al., 2008) before Nucleobond AXR 400 column purification (Machery-Nagel). Purified tRNA was hydrolysed and analysed by liquid chromatography–tandem mass spectrometry (LC–MS) (Phillips et al., 2008). For immunoblot analysis, cells grown in LB medium to an OD600 nm of 1.0 were harvested

by centrifugation, washed once in ice-cold phosphate-buffered saline and sonicated in 50 mM Tris–HCl (pH 8.0), 50 mM NaCl, 1 mM EDTA and 1 mM phenylmethylsulfonyl fluoride. Extracts were centrifuged to clear. Electrophoresis and immunoblotting were as described (Turner et al., 2005); the primary antibody was anti-pentahistidine mouse monoclonal IgG (Qiagen), dilution 1 : 1000, and the secondary antibody was goat anti-mouse IgG (H + L) alkaline phosphatase conjugate (Bio-Rad), dilution 1 : 3000. Protein was estimated by the Bradford (1976) dye-binding method using bovine serum albumin as standard. The functional importance of the eight conserved residues in the Ygf Z motif was assessed by expressing mutant YgfZ proteins from a plasmid and testing their ability to complement various phenotypes of the ΔygfZ strain.

Of note, the primer set employed for the assay with THI4 was position C (Fig. 1a) different from that (position B) for the assay for V5-tagged Pdc2p, and the pdc2Δ mutant was used instead of strain thi2Δ. We confirmed that the protein level of thi2p was also unchanged by the experimental conditions (data not shown). As expected,

V5-tagged Thi2p coimmunoprecipitated with all the THI genes except PDC5 (Fig. 1d). The association with the target DNA was also decreased by thiamin in the medium and the absence of Thi3p or, in this case, Pdc2p. These findings strongly suggest that both Pdc2p and Epigenetic screening Thi2p alone can bind target DNA and assist each other in recruitment to the THI promoters via interaction with Thi3p. We next intended to map the DNA-binding domain MAPK inhibitor in Pdc2p. Pdc2p is 925 amino acids (aa) long with an internal (aa 407–581) transactivation domain and C-terminal (aa 668–889) Thi3p-interacting domain (Nosaka et al., 2008). In addition, Pdc2p possesses putative DNA-binding

domains at the N-terminus. We constructed plasmids to produce a truncated Pdc2p with a V5-tag and used them in ChIP assays. As expected from the presumed sequence, when the N-terminal 406 aa were removed, no association with THI genes and PDC5 was detected (Fig. 1e). Conversely, V5-tagged Pdc2p(1–406) coimmunoprecipitated with all the genes tested, although very weakly as compared with intact Pdc2p (Fig. 1e). In particular, its association with THI genes was markedly reduced. The Rucaparib nmr elimination of the first ten N- or C-terminal aa from Pdc2p(1–406) led to abolishment of the ChIP signal (data not shown). Thus, the 1–406 aa region is necessary for Pdc2p to bind with target

DNA. However, this region alone may not be adequate to exert full binding activity. Alternatively, because of a lack of the Thi3p-interacting domain (Nosaka et al., 2008), it is assumed that the recruitment of Pdc2p(1–406) to the promoter regions of THI genes was decreased. Meanwhile, we attempted to locate the promoter region responsible for the expression of PDC5. Although two ethanol-repression sequence (ERA) sites are recognized in the upstream region of PDC5 (Liesen et al., 1996), it is unclear whether these cis-acting elements are involved in the induction of PDC5 gene expression in response to thiamin starvation. We constructed a series of plasmids containing terminal and internal deletions of the PDC5 promoter and used the β-galactosidase activity to monitor their promoter activities. The results are summarized in Fig. 2. The LacZ gene with PDC5′s upstream region truncated at position −418 from the start codon conferred almost full promoter activity. However, the upstream region truncated at position −390 showed significantly less promoter activity, and that at position −345 barely showed any activity. Furthermore, the deletion of the upstream region from −397 to −346 almost completely abolished the activity.

Greatest benefit from using SCR could be realised out-of-hours when GP surgeries are closed and in circumstances where information from relatives is unobtainable. This project assessed application of SCR and its impact on patient safety when used by clinical pharmacy staff working on MAU at weekends. The study was conducted over 12 weekends on an MAU at a district general hospital. Pharmacy staff working on the unit were asked to record every time SCR was accessed and whether its use resulted in an intervention; further classified into those which involved critical medicines and those with the potential to cause harm, as defined by the National Patient Safety Agency2. Data were analysed descriptively in MS Erlotinib Excel.

Ethical approval was not sought as this was a service evaluation. Over 12 weekends, 480 new patients were assessed by the pharmacy team working on MAU. Staff accessed the SCR for 183 of these patients (38.1%); information was available for 146 patients (79.8%). Of the 146 patients where the SCR was signaling pathway used, 90 patients (61.6%) had an intervention that was a direct result of having access to the SCR. This equates to 18.8% (90/480) of all patients. There were 294 interventions (average: 3.3 interventions per patient; SD 5.2; range 1 to 30). The main intervention type was regular medicines not being prescribed; 28 (9.5%) interventions involved critical medicines; 48 (16%) interventions involved patients

potentially at risk of harm if intervention had not been made. All intervention categories are detailed in Table 1. Table 1: Intervention categories for all, critical medicines, and interventions to avoid potential harm Category Number of interventions Number of interventions involving 2-hydroxyphytanoyl-CoA lyase critical medicines Number of interventions where potential harm was prevented Regular medication not prescribed 199 (67.7%) 14 (50.0%) 17 (35.4%) Allergy missing

or incorrect 45 (15.3%) 10 (35.7%) 16 (33.3%) Dose or strength incorrect 34 (11/6%) 1 (3.6%) 12 (25%) Frequency incorrect 8 (2.7%) 1 (3.6%) 2 (4.2%) Wrong medication stopped 7 (2.4%) 0 (0%) 1 (2.1%) Timing incorrect 1 (0.3%) 0 (0%) 0 (0%) Totals 294 (100.0%) 28 (100.0%) 48 (100.0%) This project has shown that one out of every five patients assessed on an MAU had an intervention that improved prescribing when the SCR was used by Pharmacy staff. In a significant minority of patients the intervention reduced potential risk of harm. For patients requiring hospital care during weekends, the SCR allows healthcare professionals to access essential clinical information that would otherwise be unavailable. Future work would include a statistical comparison against a service not using SCR during their medicines reconciliation process. 1. Greenhalgh T.,Stramer K.,Bratan T. et al. Adoption and non-adoption of a shared electronic summary record in England: a mixed-method case study. BMJ 2010; 340: 1468–5833. 2. NPSA.

, 2010). In Boyd and Linsdell’s study, excitatory rTMS (15 min of 5-Hz rTMS at 120% RMT) was applied over the left dPM before participants used their right arm to practice a continuous tracking task. The authors found that excitatory rTMS over dPM enhanced memory consolidation as indicated by off-line learning. We used a different approach by applying inhibitory rTMS (10 min of 1-Hz rTMS at 110% RMT) to dPM after practice. We found that memory consolidation, indexed

by forgetting, was impaired in those who practiced the task under the dual-task condition. However, in comparing the Control–NoTMS and Control–dPM groups, we found that rTMS applied over dPM after single-task practice did not seem to have an effect on forgetting. This INCB018424 cost suggests that the role of dPM in memory consolidation may be meditated by the practice condition. Previous studies have demonstrated

that the involvement of M1 (Kantak et al., 2010b), dorsal lateral prefrontal cortex (Kantak et al., 2010b) and supplementary motor area (Tanaka et al., 2010) in memory consolidation depends Ku-0059436 mouse on practice structure. Therefore, the engagement of dPM during memory consolidation may also depend on the structure in which the task is practiced. A key limitation of the current study is that we did not capture participants’ brain activation during practice. The selection of neural substrate hypothesised to mediate the dual task motor learning benefit was based on evidence supporting the role of Tangeritin dPM in ‘planning’ processes. A more robust design would be a combined approach in which fMRI or PET is used to identify the ‘shared neural networks’ during practice and then TMS perturbation

of the observed neural activation immediately follows practice. Second, we did not have individual brain structural MRI scans for all participants in the dPM groups. Participants without brain scans only made up a small percentage of our sample (five out of 20) and their movement times were not different from those with scans. The third limitation of the present study is that the Control–dPM group demonstrated the shortest movement time throughout the experiment (both practice and retention), even before they received rTMS application. The group had similar characteristics (age, gender) as other groups; we could not find a satisfactory explanation for this difference. Nevertheless, our primary comparison was on the measure of forgetting in which participants’ retention performance rather than practice performance was referenced to their end-of-practice performance. Thus, the difference in the Control–dPM group is not an issue for our overall conclusion. Another limitation was that we only examined the ‘planning’ processes in the present study.

, 2010). In Boyd and Linsdell’s study, excitatory rTMS (15 min of 5-Hz rTMS at 120% RMT) was applied over the left dPM before participants used their right arm to practice a continuous tracking task. The authors found that excitatory rTMS over dPM enhanced memory consolidation as indicated by off-line learning. We used a different approach by applying inhibitory rTMS (10 min of 1-Hz rTMS at 110% RMT) to dPM after practice. We found that memory consolidation, indexed

by forgetting, was impaired in those who practiced the task under the dual-task condition. However, in comparing the Control–NoTMS and Control–dPM groups, we found that rTMS applied over dPM after single-task practice did not seem to have an effect on forgetting. This BIRB 796 suggests that the role of dPM in memory consolidation may be meditated by the practice condition. Previous studies have demonstrated

that the involvement of M1 (Kantak et al., 2010b), dorsal lateral prefrontal cortex (Kantak et al., 2010b) and supplementary motor area (Tanaka et al., 2010) in memory consolidation depends selleck compound on practice structure. Therefore, the engagement of dPM during memory consolidation may also depend on the structure in which the task is practiced. A key limitation of the current study is that we did not capture participants’ brain activation during practice. The selection of neural substrate hypothesised to mediate the dual task motor learning benefit was based on evidence supporting the role of check dPM in ‘planning’ processes. A more robust design would be a combined approach in which fMRI or PET is used to identify the ‘shared neural networks’ during practice and then TMS perturbation

of the observed neural activation immediately follows practice. Second, we did not have individual brain structural MRI scans for all participants in the dPM groups. Participants without brain scans only made up a small percentage of our sample (five out of 20) and their movement times were not different from those with scans. The third limitation of the present study is that the Control–dPM group demonstrated the shortest movement time throughout the experiment (both practice and retention), even before they received rTMS application. The group had similar characteristics (age, gender) as other groups; we could not find a satisfactory explanation for this difference. Nevertheless, our primary comparison was on the measure of forgetting in which participants’ retention performance rather than practice performance was referenced to their end-of-practice performance. Thus, the difference in the Control–dPM group is not an issue for our overall conclusion. Another limitation was that we only examined the ‘planning’ processes in the present study.

All calculations were carried out using Stata 10.0 (College Station, TX, USA). A total of 101 subjects were enrolled into the study. Three participants dropped out from check details the study: one from the rifaximin group and two from the placebo group. Therefore, 98 subjects completed the study and were analyzed. Fifty-four participants were female (55%) and 44 (45%) were male, each treatment group had similar proportions of males and females (p = 0.2). The overall mean age at enrollment was 25 years (range, 18–67 y).

Thirty-six (37%) participants were enrolled during the summer months, while 62 (63%) during the fall and winter months. Eight (8%) participants were enrolled in Guadalajara and 90 (92%) in Cuernavaca. As noted in Table 1, 14 participants developed TD during the 2 weeks of study: 6 of 50 patients (12%) from the rifaximin group and 8 of 48 patients (17%) from the placebo group (p = 0.5). We also did not observe a difference GSI-IX purchase in the occurrence of MD between the rifaximin and placebo groups (p = 0.3) during the 3-week study period. We only saw a statistical difference during the first week of study for the development of MD

(p = 0.03). No difference in the occurrence of MD or TD between the two groups was seen during the second and third weeks of study. Twelve of 36 (33%) participants enrolled during the summer developed TD: 6 of 19 (32%) from the rifaximin group and 6 of 17 (35%) from the placebo group (p = 0.9). Meanwhile, 12 of 62 (19%) participants enrolled during the fall and winter developed TD: 5 of 31 (16%) from the rifaximin group and 7 of 31 (23%) from the placebo group (p = 0.5). No difference in the incidence of MD or TD was observed during each of the 3 weeks in participants treated during the summer months. We observed that participants taking rifaximin during the fall and winter months were also less likely to develop MD during the first week of study compared with those taking placebo during the same timeframe (2 of 30 [6.7%] vs 8 oxyclozanide of 29 [28%]; p = 0.04). Seventeen of the 25 participants (68%) suffering from TD provided a stool sample for microbiological analysis

before any antibiotic rescue therapy was administered: 10 (91%) from the rifaximin group and 7 (50%) from the placebo group. Bacterial diarrhea was detected in eight participants: six (60%) in the rifaximin group and two (29%) in the placebo group (p = 0.3; Table 1). Enteroaggregative Escherichia coli was the most common bacteria isolated (4 of 8), followed by enterotoxigenic E coli (3 of 8), diffusely adherent E coli (2 of 8), and Salmonella spp. (1 of 8). Two participants had mixed infections. No other bacterial or parasite pathogens were found. Three E coli isolates showed high minimum inhibitory concentration (MIC) for rifaximin (≥512 µg/mL): two of them isolated from participants in the rifaximin group and one taking placebo.

8% Indian and 6.2% others), the spectrum of diseases seen www.selleckchem.com/products/VX-809.html was as follows [disease – definite n (%), probable n (%)]: Arthritis: rheumatoid arthritis – 958 (22.9%), 68 (1.6%); osteoarthritis – 452 (10.8%), 39 (0.9%); crystal arthritis – 417 (10.0%), 18 (0.4%); spondyloarthritis – 227 (5.4%), 61 (1.5%); psoriatic arthritis – 158 (3.8%), 9 (0.2%); other inflammatory arthritis – 153 (3.7%), 94 (2.2%); Connective tissues diseases: systemic lupus erythematosus – 412 (9.9%), 26 (0.6%); vasculitis – 105 (2.5%), 22 (0.5%); Sjögren’s

syndrome – 81 (1.9%), 32 (0.8%); overlap syndromes – 73 (1.8%); scleroderma – 50 (1.2%), 4 (0.1%); undifferentiated connective tissue diseases – 45 (1.1%), 106 (2.5%); myositis – 41 (1.0%), 12 (0.3%); antiphospholipid syndrome – 22 (0.5%), 7 (0.2%); polymyalgia rheumatica – 16 (0.4%); Others: soft tissue rheumatism – 155 (3.7%); osteoporosis – 61 (1.5%); other non-rheumatologic conditions – 189 (4.5%); other rheumatologic conditions – 67 (1.6%). Rheumatoid arthritis, osteoarthritis and crystal arthritis were the three most common rheumatological diseases seen in a tertiary referral centre serving a BI 2536 supplier multi-ethnic urban Asian population in Singapore. “
“There is strong rationale for improving

care for people with chronic conditions, including osteoarthritis (OA). Successful implementation of healthcare reform requires new concepts and directions that are strongly supported by policy, new models of care (service redesign) and changes in day-to-day practice (healthcare provider and patient practice). In this paper we discuss the extent to which policy about management selleck inhibitor of OA of the hip and knee has been translated into new service models in Australia. A structured search of government and other key health websites in Australia was performed to identify policy, funding initiatives and new services models for managing OA of the hip and knee. This search

was supported by a literature review. Musculoskeletal conditions were designated a National Health Priority in Australia in 2002. Under the Better Arthritis and Osteoporosis Care initiative, Australia has developed a national policy for OA care and national evidence-based clinical practice guidelines for management of OA of the hip and knee. Only two well-described examples of new chronic disease management service models, the Osteoarthritis Clinical Pathway (OACP) model and the Osteoarthritis Hip and Knee Service (OAHKS) were identified. Primarily focused within acute care public hospital settings, these have been shown to be feasible and acceptable but have limited data on clinical impact and cost-effectiveness. While policy is extant, implementation has not been systematic and comprehensive. Clinicians have evidence-based recommendations for OA management but are poorly supported by service models to deliver these effectively and efficiently.

, 1997) using S. oneidensis MtrB as the search query. Multiple alignments of MtrB homologs were generated with clustalw (http://www.ebi.ac.uk/Tools/clustalw2/index.html)

(Chenna et al., 2003). β-Barrel architecture of the MtrB homologs was predicted selleck using the program pred-tmbb (Bagos et al., 2004). logo diagrams were generated using the clustalw alignment files (Crooks et al., 2004). mtrB was deleted from the S. oneidensis genome via application of a Shewanella in-frame gene deletion system (Burns & DiChristina, 2009). Regions corresponding to c. 750 bp upstream and downstream of mtrB were independently PCR-amplified and subsequently joined using overlap-extension PCR. Primers for mtrB deletion are listed in Table 2. The resulting fragment was cloned into suicide vector pKO2.0, which does Anti-cancer Compound Library mw not replicate in S. oneidensis. This construct (designated pKO-mtrB) was

mobilized into wild-type MR-1 via conjugal transfer from E. coli donor strain β2155 λ pir. S. oneidensis strains with the plasmid integrated into the genome were selected on solid LB medium containing gentamycin (15 μg mL−1). Single integrations were verified via PCR with primers flanking the recombination region. Plasmids were resolved from the genomes of single integrants by plating on solid LB medium containing sucrose (10% w/v) with NaCl omitted. In-frame deletions were verified by PCR and direct DNA sequencing (GeneWiz, South Plainfield, NJ). Genetic complementation of ∆mtrB was carried out by cloning wild-type mtrB into broad-host-range cloning vector pBBR1MCS (Kovach et al., 1995) and conjugally transferring the recombinant vector into ∆mtrB via biparental mating procedures (DiChristina et al., 2002). Single amino acid mutations in MtrB (C42A or C45A) were constructed using the Quickchange Lightning site-directed mutagenesis kit (Agilent Technologies, Santa Clara, CA). The mtrB gene and regions c. 750 bp upstream and downstream were PCR-amplified as a single fragment and subsequently cloned into pBBR1MCS. Mutagenesis primers C42A-sense, C42A-antisense, C45A-sense, and C45A-antisense (Table 2) were used RG7420 research buy in mutagenesis PCR

according to the manufacturer’s instructions. The resulting PCR products were subsequently transformed into XL10 Gold KanR competent cells (Agilent Technologies). Correct amino acid mutations (C42A or C45A) were verified by direct DNA sequencing using primers MTRB-SeqF and MTRB-SeqR (Table 2). The mutated mtrB constructs were subsequently cloned into suicide vector pKO2.0 and were ‘knocked in’ to the native chromosomal position. Nucleotide sequence changes were verified by PCR and DNA sequencing of S. oneidensis ‘knock-in’ transformants. Genetic complementation of mutant C42A was carried out by cloning wild-type mtrB into broad-host-range cloning vector pBBR1MCS (Kovach et al., 1995) and conjugally transferring the recombinant vector into mutant C42A via biparental mating procedures (DiChristina et al., 2002).