digitatum Pd01 was performed using a Roche GS-FLX sequencer with a half plate run. A total of 519 480 reads were finally obtained and automatically assembled by newbler software (454 Life Science; Li HY et al., unpublished data). All the assembled contigs were aligned to the mitochondrial genome of P. chrysogenum by BLASTN (Altschul et al., 1997), and contigs showing high identity were screened out as fragments of Pd01 mitochondrial DNA. PCR amplification was performed to finish and verify the mitochondrial genome sequence. The nucleotide sequence of AG-14699 the P. digitatum Pd01 mitochondrial genome has been deposited in GenBank under accession number HQ622809. Coding regions of the mitochondrial genome

were predicted by glimmer3.0 and manually checked (Delcher et al., 1999). Introns and rRNA genes were mainly identified by blast pairwise comparison between mitochondrial genomes of P. digitatum, Penicillium marneffei and P. chrysogenum. tRNA genes were predicted by tRNAscan-SE using the genetic code of mould (Lowe & Eddy, 1997). Cumulative codon usage and amino acid usage were calculated by gcua software (McInerney, 1998). Maximum-parsimony analysis was carried out with the DZNeP concatenated sequences of 14 mitochondrion encoded proteins (cox1, atp9, nad3, cox2,

nad4L, nad5, nad2, cytb, nad1, nad4, atp8, atp6, nad6 and cox3) using mega 4 software (Tamura et al., 2007). Sequence data used in comparative analysis and phylogenetic tree construction were obtained from GenBank: Allomyces macrogynus (NC_001715), Cryptococcus neoformans var. grubii (NC_004336), Saccharomyces cerevisiae (NC_001224), P. marneffei (NC_005256) and P. chrysogenum (AM920464). The draft second assembled genome of P. digitatum Pd01 consists of about 11 000 contigs, and one of them was predicted as a mitochondrial DNA fragment based on its high similarity to that of P. chrysogenum (>95%). A total of 10 026 reads were clustered

into this contig with the average coverage being 134.6-fold, while the average coverage of the chromosome was 12.5-fold. Subsequent PCR amplification was carried out and the mitochondrial genome was completed by sequencing of the products. The mitochondrial genome of P. digitatum Pd01 is a circular molecule with a length of 28 978 bp and an average A + T content of 74.7%. Fifteen protein coding genes were identified, as well as 27 tRNA genes and the large and small subunits of ribosomal RNA (rnl and rns), accounting for approximately 80% of the whole mitochondrial genome in total. All fifteen mitochondrial protein coding genes were conserved between P. digitatum, P. marneffei and P. chrysogenum, while levels of amino acid pairwise sequence similarity varied between 73% and 100% (Table 1). The rps5 gene was the least homologous followed by nad6. Other genes showed more than 90% amino acid identity between the three species, and atp8 was the most conserved, showing 100% identity between the three Penicillium species.

Quantitative invasion assay values were calculated as follows: We used an in vitro assay according to Trombert et al. (2010), modified from the method described by McCormick et al. (1993). Briefly, the colon carcinoma HT-29 cell line was grown to confluence (18–21 days) on 3.0-μm Belnacasan cost pore-size filters (or transwells, Millicell®, Millipore) with glucose-free RPMI (Gibco). Each transwell was inoculated individually to the

apical surface with 400 μL of approximately 1 × 107 CFU mL−1 of bacterial cultures and immediately incubated for 60 min at 37 °C. After extensive washing with sterile PBS, the extracellular bacteria were killed by treatment of monolayers with gentamicin (50 μg mL−1). Immediately after gentamicin treatment, the medium from basal compartment

of the epithelial cell monolayer was collected and plated for CFU to assess the number of bacteria that passed through the cell monolayer. The polarization of cells was confirmed by transepithelial electrical Pifithrin-�� in vitro resistance and transmission electron microscopy (data not shown). All results are expressed as means±SD of an individual experiment performed in triplicate. P-values were calculated according to Student’s t-test, and P<0.05 or P<0.01 values were considered statistically significant. To assess whether the sopD2 locus is a pseudogene in the serovar Typhi, we compared the available sequences of S. Typhi CT18 and S. Typhi Ty2 (McClelland et al., 2001; Parkhill et al., 2001; Deng et al., 2003). We observed that the sequence of sopD2 in S. Typhi contains a frameshift at codon 48 resulting in

a premature stop codon that disrupts the expected ORF. Accordingly, the online databases report sopD2 as a pseudogene (Fig. 1). To confirm the presence of this frameshift in our S. Typhi clinical strain collection, PCR assays were carried out using SopD21 and SopD22 primer pairs. The primers yield an 1800-bp amplicon in both S. Typhi and S. Typhimurium and were used to test the clinical strains obtained from Chilean typhoid patients (STH collection, Hospital Dr Lucio Córdova, Chile). The PCR products were sequenced and in silico comparison was performed with the reference strains (S. Typhi CT18, S. Typhimurium LT2 and Salmonella enterica serovar Paratyphi A ATCC 9150) using C59 clinical trial the bioinformatic available program (clustalw; http://www.ebi.ac.uk/Tools/msa/clustalw2/). Our results indicated that the deletion described in S. Typhi CT18 is conserved among S. Typhi clinical strains (Fig. 2). Therefore, the sopD2 frameshift mutation seems to be a feature in the genome of serovar Typhi. Recently, we reported that S. Typhi harboring the sseJ gene from S. Typhimurium significantly disrupted HT-29 monolayer compared with the wild-type strain (Trombert et al., 2010). In the same way, we infected polarized HT-29 monolayers with the S. Typhi wild-type and S. Typhi sopD2STM strains.

, 1995). IF3 is known to promote subunit dissociation and to discriminate other tRNAs from the initiator tRNA; mutations at positions 787, 791, 792, and 795

result in a decrease in subunit association (Tapprich et al., 1989; Santer et al., 1990; Lee et al., 1997). The decreased binding affinity of IF3 to 30S when residues at 791 and 792 are altered (Tapprich et al., 1989; Santer et al., 1990) indicates that this is not due to a lack of antiassociation activity, but rather a loss of ability of the initiator tRNA to select or bind to the P-site, resulting in a decrease in subunit association. The footprinting and structural data suggest that ABT-199 in vitro these residues are heavily involved in tRNA selection in the P-site and it is likely that these sites may form a structural motif that interacts with IF3 and recruits the initiator buy Dorsomorphin tRNA to the P-site. The P-site-specific antibiotics edeine, pactamycin, and kasugamycin, which stabilize

the P-site-bound tRNA, show a footprint in the 790 loop; this also supports the involvement of the 790 loop in the recruitment of initiator tRNA to the P-site. Here, we describe how we functionally analyzed the role of G791 in protein synthesis. This residue has been shown to be an invariant and essential residue for ribosome function (Lee et al., 1997; Song et al., 2007). To investigate the functional role played by G791 during the process of protein synthesis, we adopted a novel genetic approach (Lee et al., 1997, 2001; Kim et al., 2009) by introducing a base substitution at position 791 and then selecting multicopy suppressors that partially restored the protein synthesis ability of the mutant ribosomes. We identified IF1 as a multicopy suppressor of the mutant ribosome bearing a G to U substitution at position 791. Based on functional analyses of the effects of IF1 on the mutant ribosome, we suggest the involvement Phosphatidylinositol diacylglycerol-lyase of IF1 in the restoration of the P-site that

was perturbed by a nucleotide substitution at position 791. All plasmids were maintained and expressed in E. coli DH5α (Hanahan, 1983). Cultures were grown in Luria–Bertani (LB) medium (Luria & Burrous, 1957) or LB medium containing 100 μg mL−1 ampicillin (LB–Amp100). To induce the synthesis of plasmid-derived rRNA from the lacUV5 promoter, IPTG was added to a final concentration of 1 mM and 0.1% of l-arabinose was used to induce the synthesis of initiation factors from the BAD promoter. Plasmids pRNA122 and pRNA16 ST were described previously (Lee et al., 1997, 2001). The construction of pKAN3, pKAN4, and pKAN6 was described previously (Tamura et al., 2006). To construct pRNA122-U791U1192 and pRNA122-G770U1192, the XbaI and SexAI fragment from pRNA122-U1192 (Lee et al., 1996) was subcloned into the same sites in pRNA122-U791 (Song et al., 2007) and pRNA122-G770 (Kim et al., 2007).

The activities of the other σH-dependent promoters preceding sigN (PN1) and rpp operon (Prpp), which are also known for their dependence on σH (Takano et al., 2007),

were also significantly downregulated in the bldG mutant (Fig. 3a). These observations supported the hypothesis that BldG regulates the activity of σH and alternative sigma factors by binding to RshA. Further, the σH-dependent transcription was studied by an in vitro transcription assay (Fig. 3b). As described in previous studies, RshA inhibited the σH-dependent transcription at PH1. This RshA-dependent transcriptional repression was abolished in a dose-dependent manner by the addition of BldG at excess molar ratios (Fig. 3b). This finding can be attributed to the dissociation of σH from RshA, which in turn

binds to BldG. The lines of evidence obtained in this study suggest that the role of BldG is highly selleck inhibitor pleiotropic. BldG regulates the expression of both developmental and stress-responsive genes in S. griseus. Since σH and its paralogs are not essential for the initiation of development (Takano et al., 2007), BldG probably binds to another sigma-factor antagonist involved in the developmental control. Recently, Parashar et al. (2009) reported that BldG binds NVP-LDE225 molecular weight to the putative anti-sigma factor encoded by SCO3548, the adjacent cds, to control the key developmental processes in S. coelicolor A3(2). The specific sigma factor regulated by this anti-sigma factor is expected to be involved in developmental control, although this sigma factor has not yet been identified. The conserved gene organization suggests that these findings would also be observed in S. griseus. During the writing of this

paper, Sevcikova et al. (2010) reported a similar observation on the interaction between BldG and RshA in S. coelicolor A3(2). The authors demonstrated specific interaction between BldG and UshX (the RshA ortholog) by pull-down and two-hybrid analyses and showed that the activity of the σH-dependent promoter preceding ushX-sigH operon (sigHp2; equivalent of PH1 of S. griseus) is abolished in a bldG mutant of S. coelicolor. Overall, our results are confirmatory except that the activities of the σH-independent promoters preceding rshA (PH2 and PH3) were reduced in the bldG mutant of S. griseus (Fig. 3a). In contrast, the corresponding promoters Adenosine triphosphate of S. coelicolor (sigHp3 and sigHp4) were upregulated in the bldG mutant of S. coelicolor A3(2) (Sevcikova et al., 2010). Currently, we do not know why the σH-independent promoters were also affected by the knockout of bldG, but the difference in the two species implies that this is due to some indirect effect. On the other hand, the identical evidence regarding σH-dependent promoters obtained in the two phylogenetically divergent species strongly suggests that the regulation generally occurs in this group of organisms. Similarly as in S.

“
“UK guidelines recommend routine HIV testing in general clinical settings when the local HIV prevalence is > 0.2%. During pilot programmes evaluating the guidelines, we used laboratory-based testing of oral fluid from patients accepting tests. Samples (n = 3721) were tested

manually using the Bio-Rad Genscreen Ultra HIV Ag-Ab test (Bio-Rad Laboratories Ltd, Hemel Hempstead, UK). This was a methodologically robust method, but handling of samples was labour intensive. We performed a validation study to ascertain whether automation of oral fluid HIV testing using the fourth-generation HIV test on the Abbott Architect (Abbott Diagnostics, Maidenhead, UK) platform was possible. Oral fluid was collected from 143 patients (56 Selleckchem Protease Inhibitor Library known HIV-positive volunteers and 87 others having contemporaneous HIV serological tests) using the Oracol+ device (Malvern Medicals, Worcester, UK). Samples were tested concurrently: manually using the Genscreen Ultra test and automatically on the Abbott Architect. For oral fluid, the level check details of agreement of results between the platforms was 100%. All results

agreed with HIV serology. The use of the Oracol+ device produced high-quality samples. Subsequent field use of the test has shown a specificity of 99.97% after nearly 3000 tests. Laboratory-based HIV testing of oral fluid requires less training of local staff, with fewer demands on clinical time and space than near-patient testing. It is acceptable to patients. The validation exercise and subsequent clinical experience

support automation, Tobramycin with test performance preserved. Automation reduces laboratory workload and speeds up the release of results. Automated oral fluid testing is thus a viable option for large-scale HIV screening programmes. Since 2007, a change in the HIV testing paradigm in the UK has been proposed to reduce both undiagnosed and late-stage diagnosed HIV infection. Guidance from the National Institute for Health and Clinical Excellence follows that from the British Association for Sexual Health and HIV, and the British HIV Association, in calling for more widespread testing, including routine HIV testing in general medical settings in areas where HIV prevalence exceeds 0.2% [1-4]. Expansion of HIV testing has driven the development and appraisal of new HIV testing technologies, such as near-patient point-of-care tests (POCTs) and the use of various biological specimens to diagnose HIV infection, including whole blood, serum, capillary blood, dried blood spots and oral fluid. Oral fluid testing has several advantages over blood-based techniques: it is less invasive and less painful, the specimen collection can be performed by the patient without direct supervision, and oral fluid sampling is likely to be less hazardous to health care personnel. To date, the only licensed oral fluid-based HIV test is the OraQuick® ADVANCE Rapid HIV-1/2 Antibody test (OraSure Technologies, Inc.

After electrophoresis, gel was stained with Coomassie Brilliant Blue R-250. For activity staining, zymographic analysis of the protease was performed using gelatin (0.1%) as the substrate as Selleckchem Roxadustat described by Karbalaei-Heidari et al. (2009). Zymographic analysis for the amylase was performed on nondenaturing electrophoresis slab gels (10% polyacrylamide) prepared with 10% of sucrose, as described by Cadenas & Engel (1994). The amylase activity, with soluble starch as the substrate,

was determined using DNS (3,5-dinitrosalicylic acid) method (Miller, 1959). One unit (U) of amylase activity was defined as the amount of enzyme necessary to produce 1 μmol of reducing sugar per minute under the assay conditions. Protease activity was measured as described previously (Karbalaei-Heidari et al., 2009). One unit (U) of protease activity was defined as the amount of enzyme yielding 1 μmol of tyrosine per minute under the assay conditions. The effect of pH on enzyme activity was studied over a pH range of 4.0–12.0. The pH stability of the enzymes was determined by incubation with different buffer systems at 30 °C for 24 h. The following buffer systems (100 mM) were used: glycine-HCl buffer, pH 4.0; sodium acetate buffer, pH 5.0–6.0; potassium phosphate buffer, pH 7.0; Tris–HCl buffer, pH 8.0–8.5; glycine–NaOH buffer, pH 9.0–12.0. To investigate the effect of temperature, the assay

was conducted under different temperatures from 30 to 90 °C. The thermostability of the enzyme was determined by pre-incubating CP-868596 order the enzyme sample at various temperatures for 24 h, and residual activity was measured Selleckchem Ixazomib using the standard assay. The activity of the purified enzyme was measured in enzyme reaction mixture containing 0–20% NaCl. Salt stability of the enzyme was determined by incubating

the enzyme with different concentrations of NaCl for 24 h, and the remaining activity was determined under standard assay conditions. The effect of organic solvents with different log Pow values at 50% (v/v) concentration on the purified enzyme was determined by incubating the enzyme solution in different organic solvents at 30 °C. Residual activity was measured under the standard conditions. If residual activity was more than 50% after 10 days, half-life was taken as ‘> 10 days’. While activity was < 50% after 1 h, half-life was taken as ‘< 1 h’. Effects of different metal ions and chemical reagents [ethylenediaminetetraacetic acid (EDTA), phenylmethylsulfonyl fluoride (PMSF), phenylarsine oxide (PAO), diethyl pyrocarbonate (DEPC), β-mercaptoethanol, SDS, Triton X-100, and Tween-80] on the activity of purified enzymes were examined after they had been pre-incubated with them at 30 °C for 12 h, respectively, and the residual activity was determined under optimal assay conditions. Activity of the enzyme assayed in the absence of any additives was taken as 100%.

Grading: 1D Where a woman chooses to breastfeed against the medical advice in Recommendation 8.4.2, she and the baby should be monitored regularly for maternal adherence to ART; VL monitoring of the mother and diagnostic testing of the baby should be performed regularly (monthly). If the mother’s NVP-BGJ398 datasheet adherence is suboptimal

or she has detectable viraemia or an intercurrent illness that affects her ability to take or absorb ART, or she develops mastitis, she should be advised again to stop breastfeeding. 8.4.5 All infants born to mothers infected with HIV should have an antibody test at age 18 months. Grading: 1C The potential for breastfeeding emphasizes the possibility of late transmission of HIV after the standard 3-month PCR test. Babies known to be breastfed should be tested monthly by PCR as above, but not all breastfeeding will be disclosed, and all babies born to HIV-positive women should have a negative HIV antibody test documented at age 18 months

(see Section 8.5: Infant testing below). 8.5.1 HIV DNA PCR (or HIV RNA testing) should be performed on the following occasions (Grading: 1C): During the first 48 h and before hospital discharge. LGK-974 price 2 weeks post infant prophylaxis (6 weeks of age). 2 months post infant prophylaxis (12 weeks of age). On other occasions if additional risk (e.g. breastfeeding). HIV antibody testing for seroreversion should be checked at age 18 months. The gold standard test for HIV infection in infancy was HIV DNA PCR on peripheral blood lymphocytes, although a number of studies, including the large French perinatal cohort have now demonstrated equal or increased early sensitivity with amplification of viral RNA with no false positives [71]. Infants infected intrapartum may have low

peripheral blood HIV levels, so HIV DNA/RNA may not be amplified from all infected infants at birth. Indeed a positive HIV DNA PCR result within 72 h of birth is taken as presumptive evidence of intrauterine transmission. Within Branched chain aminotransferase the first few weeks of life, sensitivity of the viral diagnostic tests increases dramatically and by 3 months of age, 100% of non-breastfed HIV-positive infants are likely to be detected [72]. In view of the genomic diversity of HIV where infant diagnosis will rely on HIV DNA amplification, a maternal sample should always be obtained for HIV DNA amplification with, or prior to, the first infant sample to confirm that the primers used detect the maternal virus. If the maternal virus cannot be detected then a different primer set and/or test should be used. Infant HIV diagnostic testing should be undertaken at birth, 6 weeks and 12 weeks of age. Evidence from the French perinatal cohort demonstrated that neonatal ART, especially if more than one drug, can delay the detection of both HIV DNA and RNA in the infant [73].

Furthermore, the quality and robustness of study designs were not assessed in this review as the main focus was to extract data regarding study methods and the inclusion or exclusion of performance feedback. Future studies, therefore, may wish to include assessment of study design quality. Eleven studies used the simulated-patient method in a purely covert manner, as a tool for assessing practice behaviour of pharmacists and their staff, although a further 10 did not provide immediate feedback, hence also using the study mainly for performance assessment. learn more This finding is in accordance with a recent systematic review by Mesquita et al., whereby the results showed that

the focus of simulated-patient methods was primarily on assessment, rather than as an educational focus for enhancing practice skills of pharmacists and their staff.[19] This also highlights that although simulated patients have been used in pharmacy practice, little published

information and regard has focused on the role of performance feedback and training in pharmacy education, in shaping the behaviour of pharmacists and their staff.[19,45] The literature has revealed that this method may be a valuable tool to shape practice behaviour and, in the few studies that monitored acceptance of this as an educational tool, participants rated the experience positively. Therefore, because of its face validity, reliability and acceptance, it could be more widely used as a tool to assess current check details training needs in community pharmacy, identify potential barriers to change, and to compare different practice change strategies.[12,15,20] Of course to be used for educational purposes, i.e. to improve performance through immediate feedback, the simulated-patient visits need to be conducted with prior consent from participants to involve the Hawthorne effect, whereby performance improves with the

knowledge of impending assessment.[1,11] The Hawthorne effect is desirable, as it enables pharmacists and their staff to maintain a high level of performance, which then becomes routine practice. Therefore, simulated patients used in a consented, educational and Farnesyltransferase reflective framework, as part of continuing professional development, aims to improve future performance.[36] Written notes or checklists, documented as soon as possible after simulated-patient visits, were the most common method of data collection. This was also found by a systematic review by Watson et al.[23] In fact, the use of self-completed questionnaires are the most common method of data collection in pharmacy practice research in general.[28] However, problems can arise as a result of time separation between observation and data recording, with regards to the fallibility of recall and memory.

Retrospective analysis of patient records over a 24-month period, looking at CBCT examinations performed on subjects under 18 years of age. Clinical indications, region of interest, scan field of view (FoV), incidental findings and exposure factors used were recorded. There were 294 CBCT examinations performed in this age group, representing 13.7% of all scanned patients. CBCT was used more frequently in the >13 year age group. The most common use was for localisation of unerupted teeth in the anterior maxilla BMN 673 datasheet and the detection of root resorption.

Optimisation of X-ray exposures did not appear to be consistent. When planning a CBCT service for children and young people, a limited FoV machine would be the appropriate choice for the majority of clinical requirements. It would facilitate clinical evaluation of scans, would limit the number of incidental findings and contribute to optimisation of radiation doses. “
“Children and adolescents with cystic fibrosis (CF) are believed to be at low risk for dental caries, but this paradigm has not been critically mTOR inhibitor evaluated. To conduct a qualitative systematic review of the international literature on dental caries prevalence in children and adolescents with CF and make recommendations on future CF-related oral health research priorities. The Preferred Reporting Items for Systematic reviews and Meta-Analyses

(PRISMA) statement was used to identify relevant studies published between 1960 and 2013. The search resulted

in 696 studies. Fifteen publications were included in the qualitative systematic review. Ten studies concluded that children with CF had significantly lower caries prevalence than control children, three studies reported that children with CF had higher caries prevalence, and two studies found no difference by CF status. Of the seven studies including age-based subgroup analyses, only one study supported the current paradigm. All studies had limitations that may bias study results. While children with CF may be a lower risk for dental caries, adolescents with CF may not be at lower caries than those mafosfamide without CF. Additional research is needed to evaluate a potentially flawed paradigm regarding caries risk in children and adolescents with CF. “
“International Journal of Paediatric Dentistry 2011; 21: 217–222 Background.  More than one-quarter of New Zealand children are overweight or obese. Research on the causes of obesity has found associations with high consumption of sweetened foods and beverages, which have also been shown to be risk factors for dental caries, but studies investigating a possible association between dental caries and obesity have had conflicting findings. Aim.  The aim of this study was to determine whether deciduous dental caries experience was associated with BMI among paediatric dental clinic attenders. Design.

Substitution of two conserved residues (G49 and L107) from MtbPDF with the corresponding residues found in human PDF affected its deformylase activity. Among characterized PDFs, glycine (G151) in motif III instead of conserved aspartate is characteristic Avasimibe of M. tuberculosis. Although the G151D

mutation in MtbPDF increased its deformylase activity and thermostability, it also affected enzyme stability towards H2O2. Molecular dynamics and docking results confirmed improved substrate binding and catalysis for the G151D mutant and the study provides another possible molecular basis for the stability of MtbPDF against oxidizing agents. Proteins evolve by rare mutations that provide functional innovations without affecting the pre-existing global structure and activity (Bowie et al., 1990). As beneficial mutations are rare, the ability of an enzyme to accumulate sequence changes and maintain the required activity for better survival of the host organism is an important aspect of its evolvability (Woycechowsky et al., 2008). The emergence of multiple drug-resistant strains of Mycobacterium tuberculosis, a synergy between HIV and M. tuberculosis infection, and a need for shortened chemotherapy for tuberculosis treatment have increased the demand for improved drugs with alternative targets. Peptide

deformylase (PDF; EC 3.5.1.31), encoded by the def Selleck Venetoclax gene, catalyses the removal of the formyl group from N-terminal methionine following translation. This enzyme, present in all eubacteria and in eukaryotic organelles, is a potential target for discovery of antibacterial agents (Guay, 2007). Its essentiality for survival has been demonstrated for many bacteria, including Mycobacterium bovis (Teo et al., 2006). Most of the PDF inhibitors available are derivatives of the natural deformylase inhibitor actinonin, and many, such as LBM-415, have progressed to preclinical

and clinical stages of development (Chen et al., 2000; Butler & Buss, 2006). However, the published structural evidence for similar binding of actinonin to human PDF has complicated the whole drug discovery process based on PDF (Escobar-Avarez et al., 2009). Thus, the available sequence variations between bacterial and human PDFs need old to be explored further to identify structural variations between the two for designing novel PDF inhibitors. Characterizing the amino acid sequence variations between the PDF of M. tuberculosis (MtbPDF) and other PDFs might help us to design specific inhibitors targeting MtbPDF. Here recombinant MtbPDF and its selected substitution mutants were characterized to study the properties of this enzyme and to define the role of substituted residues in its activity and stability. All the routine chemicals, reagents, substrates, culture media and antibiotics were purchased from Sigma-Aldrich. PCR primers were obtained from Integrated DNA Technologies. Mycobacterium tuberculosis H37Rv genomic DNA was obtained from Colorado State University.