Effects Proteomic Examination of HT 29 colon cancer cells taken care of with RSV and IGF 1 HT 29 cells were handled with either DMSO, IGF 1 or RSV. LC/MS/MS Mud PIT evaluation of HT 29 cells detected 1231 proteins amongst all three treatment method groups. Using quantile nor malization strategy allowed us to scale all protein abundances in all samples for the similar distribution. This permits reasonable estimation of fold modify between experimental situations. The listing of recognized proteins and respective normalized fold adjustments were imported into GeneGo Metacore soft ware to analyze cellular processes considerably altered by IGF 1 and RSV with respect to control. Fifty most dif ferentially represented biological pathways had been identi fied from the set of proteins across all experimental disorders and are presented in Figure S2.
Big pathways impacted by RSV and IGF one consist of these responsible for cytoskeleton remo deling, apoptosis signaling, cell cycle, cell adhesion, inflammation and glucose metabolic process. Pathways that had p 0. 05 have been thought of important. RSV responsive pathways, the PPP and the cytoskeleton remodeling pathway, which play a essential role in cancer cell kinetics, were selected to determine I-BET151 Histone Methyltransferase inhibitor if RSV suppression of proliferation and induction of apoptosis involves suppression of metabolic pathways such since the PPP and disruption of cell ECM crosstalk. IGF one and RSV modulate the PPP to target cell cycle progression in human colon cancer cell line HT29 PPP is often up regulated in cancer cells.
Proteomic evaluation unveiled that G6PDH and TKT the two critical enzymes you can check here from the oxidative and non oxidative branches of your PPP respectively, were altered by RSV or IGF one treatment. We observed that RSV attenuated G6PDH and TKT levels. Outcomes employing enzyme kinetic assays for both G6PDH and TKT assistance these observa tions. RSV suppressed and IGF 1 elevated each G6PDH and TKT ranges. Even while in the presence of IGF one, RSV suppressed G6PDH and TKT enzymatic levels confirming its efficacy in obese situations. Potentiated G6PDH and TKT suppression by RSV while in the presence of IGF one may very well be because of up regulation of reactive oxygen species by both IGF one and RSV at large concentration leading to elevated apoptosis. RSV suppresses even though IGF one elevates the talin Fak signaling pathway Proteomic analysis unveiled that the cytoskeleton remo deling pathway is among the vital pathways altered within the HT 29 colon cancer cells handled with RSV or IGF 1.
As cytoskeletal components make up a significant portion of complete protein in most tis sues, it’s not surprising to view quantitative modifications within this category of proteins. Among the list of significant signaling pathways during the cytoskeleton remodeling, is definitely the interac tion among proteins talin and FAK. Both RSV and IGF one altered the talin FAK signaling pathway, but the differences weren’t sizeable.
The whole exome data in the 18 lung tumors and matched typical samples had been captured implementing Agilent SureSelect 38 M kit and sequenced on an Illumina HiSeq 2000 platform as published. On typical, 48 Mb paired finish reads were garnered per sample with an typical sequencing depth of 63? in target regions. Overall, the sequence reads covered 98. 9% bases with the target areas by not less than one particular read through and 79. 9% bases by a depth of at the least 20?. SAMtools was applied to characterize sSNVs in these samples, Sanger sequencing was utilized to validate functionally essential sSNVs. We’ve got produced the unique sequence information on the market at. We also incorporated NGS data from 7 lung cancer cell lines. Two of them, Pc 9/S2 and Pc 9/BRc1, have been sequenced on an Illumina HiSeq 2000 platform applying Nimblegen SeqCap Ez Exome Library kit v2.
We obtained 8. four ? 109 bases of quick reads for Pc 9/S2 with an typical of 232. six? coverage, and seven. eight ? 109 bases of brief reads for Computer 9/BRc1. selleck The other five cell lines, HCC827, HCC827/R1, HCC827/R2, HCC4006, and HCC4006/ER, have been sequenced on an Illumina HiSeq 2000 platform employing the Agilent SureSelect 38 Mb kit for full exome sequencing. Their common coverage is all 109?. For these 7 cell lines, the sequence reads covered 98. 9% bases from the target areas by not less than one particular read and 85. 5% bases by a depth of at least 20?. Eight pairs of cell lines have been in comparison to identify sSNVs that were special to drug sensitivity or drug resistance cell lines. Especially, the somatic model was executed by designating the targeted cell line as tumor as well as cell line to become compared as nor mal.
The sSNVs that resulted in the examination have been then Naftopidil experimentally validated by Sanger resequencing. Cell line DNAs were employed as template for PCR amplifi cation. M13 tagged gene exact primers were made working with Primer 3 program. Sequence chromatograms were analyzed utilizing Mutation Surveyor software and manual inspection. The information is usually noticed while in the unique do the job. We also simulated WES of 10 tumor normal pairs utilizing the profile based Illumina pair finish Read through Simulator. Our simulation method and corresponding command lines had been described in detail in Additional file 2. We fixed the insert size of the simulated reads at 200 bp. The read length and typical coverage were set to 75 bp and a hundred?, respectively. In addition, we allow the frequency of sSNVs in every single sample be 10 occasions larger than that of indels and structural variants be 10 occasions less than indels. Mainly because tumor samples carry driver mutations, we allow the frequency of SNVs inside the tumor be increased than that within the normal sample. Alignment We utilized BWA to align brief sequencing reads to the UCSC human reference genome hg19. The de fault arguments of BWA had been applied for the alignment.
But, we observed slow costs of HA H3. three replacement at telomeres outside S phase, indicating slow replication independent histone exchange that may be necessary for upkeep of telomeric stability. It can be identified that repetitive features of telomeres and pericentromeres possess a rather very low affinity for nucleosomes and this may contribute to his tone displacement. Discussion Nucleosome dynamics and histone turnover are certainly not very well understood in mammals. In this report, we’ve devel oped a robust, TET inducible procedure to review the kinetics of genome wide deposition within the histone variant H3. 3 in MEF cells. Applying this program, we have been in a position to measure turnover prices of genome wide H3. three target websites and were ready to infer essential variations between turnover rates of many genomic areas.
Numerous categories of H3. three deposition kinetics were observed, invariably large turn above at promoters, a broad range of turnover prices at enhancers, and slow turnover at gene bodies. In addition, repeat aspects displayed radically various turnover rates with reasonably higher turnover selleck at tRNA and rRNA ele ments and quite slow H3. three exchange at telomeres. These benefits deliver novel insights into the genome wide flip above of H3. three containing nucleosomes and suggest distinct mechanisms of nucleosome assembly, stability and eviction so as to fulfill their function in regulating transcription and keeping chromatin stability and integrity. Genome wide incorporation of H3. 3 Contrary to canonical histones, the histone variant H3. three is usually deposited onto chromatin inside a replication independent manner.
It really is enriched in transcriptionally active re gions, such as gene bodies and promoters and enhancers of mammalian cells. Incorporation of H3. 3 into chromatin destabilizes the nucleosome framework and may facilitate transcriptional activation by creating a much more available chromatin configuration. Its de position selleck chemical at promoters also as in gene bodies is asso ciated with epigenetic inheritance and could contribute to epigenetic memories. 48 hrs and reached a maximum at 72 hrs right after induction, suggesting that a distinct mech anism for the HA H3. three deposition could take impact at telomeres. Heterochromatic areas are expected to possess Our analysis of H3. three distribution in the regular state revealed its presence at intergenic regions that overlap with histone modifications that often mark enhancers, such has H3K4me1, H3K27ac and H2A.Z, in addition to the enrichment at TSS areas. In coding regions, we find that H3. 3 is broadly distributed inside the gene physique of actively transcribed genes likewise as TESs and regions straight away just after. This association is very well corre lated with gene expression levels as has been reported from other cell lines.
Numerous studies have established that whey protein could possibly exert sa tiating results and minimize adiposity in rats. In explaining this impact, authors through the later examine propose that whey derived proteins do elicit a satiating impact with the enhanced secretion of gut neuropep tides such as cholecystokinin or glucagon like peptide 1. Thus, this effect might possibly have already been observed in our research while examining circulating CCK and GLP 1 was past the scope of our investiga tion. With regard to entire body composition alterations, how ever, the feeding intervention in our examine didn’t confer modifications in body unwanted fat from the protein supplemented disorders. Likewise, the feeding intervention did not in crease DXA lean entire body mass which is demon strated from the aforementioned rodent research that chronically fed rats whey protein above a 25 day time period.
However, that Pichon et al. implemented dissection approaches to assess physique composition whereas our DEXA approach could introduce a larger degree of error which selleck chemical could have obscured our findings. Furthermore, we can not rule out the hypothesis that consuming higher professional tein diets over longer periods minimizes adiposity and enhances and/or major tains muscle mass throughout maturation and subsequent aging in people, respectively. It really is also noteworthy mentioning that one can find limita tions on the existing research. Initially, rodents had been examined as opposed to people with regards to learning leucine, in sulin, and toxicological responses to these whey protein sources.
It must be noted, nevertheless, that rats and people seem to reply similarly to whey protein as it continues to be shown to boost circulating leucine and mar kers of muscle Ki16425 protein synthesis following exercise in both species. Consequently, we hypothesize that human responses will probably be equivalent when examining the physiological effects of WPH versus WPI dietary supplements. With regard on the current toxicology research, it should really be noted that only five animals were examined per situation above a thirty day feeding time period. In parallel to our study, nonetheless, you will find other latest studies examining the toxicological effects of other compounds which have similarly studied six animals per situation. Creat ine monohydrate is also a serious ingredient while in the WPH based mostly supple ment. Having said that, creatine monohydrate isn’t going to alter glucose tolerance or insulin sensitivity and it is not insuli nogenic nor does it have an effect on circulating leucine concentra tions. With regard to other main elements present inside the WPH based supplement, L citrulline hasn’t been proven to affect circulating insulin and/or leu cine ranges, although vitamin C continues to be shown to cut back insulin in variety II diabetes individuals over persistent supplementation intervals, and L lysine could possibly stimu late insulin secretion from pancreatic beta cells.
DNA extraction efficiency and good quality from biogas samples have also been compared to PCR primarily based analyses, but a robust strategy, especially for analyzing samples from SW biogas fermentation materials and based on substantial throughput shotgun pyrosequencing, has yet to be reported. Toward this end, we 1st evaluated five DNA extraction protocols for samples collected from a mesophilic SS AD fermenter fed with SW. Immediately after the T RFLP evaluation, we then chose the two considerably better protocols and ready DNA samples for our pyrosequencing based metagenomic research. Our effects have led to novel insights into microorganism composition, gene content material, and metabolic capacity with the SW fermentation. Success and discussion Evaluation of DNA extraction techniques for large throughput pyrosequencing Biogas fermentation samples are very complex because of the presence of a variety of natural compounds and various degradation products.
In SS AD samples, microorganisms bind strongly to strong supplies and have selleck chemical VEGFR Inhibitors a rather heteroge neous distribution within the samples. To be able to discover a far better protocol to the isolation of large quality DNA preparations for pyrosequencing, we set out to assess five DNA extraction solutions. Applying electrophoresis assay for checking quality and yield of genomic DNA extracts, we identified that Protocols E, EY, and F gave rise on the highest yields, ranging from 160. 5 ngul to 121. four ngul, whereas Protocol P made the lowest yield, with 20. five ngul. Yet, Protocol F showed the highest degree of smearing and each Protocols F and S showed very low purity based on A260A230 ratios.
The DNA extract from Protocol S appeared dark yellowish and its high quality couldn’t be measured primarily based on spectrophotometry. The distinctions kinase inhibitor ABT-263 between the 5 solutions were mostly observed on the cell lysing actions, which are vital for DNA yield and excellent primarily when discipline sampling could be the only source. According to our outcomes, Protocol P showed the lowest DNA yield, suggesting inadequate lysing regardless of using vigorous mechanical force, especially when in contrast with all the corresponding procedures in other linked protocols. Thus, we recognized that furthermore to mechanical forces, lysis reagents employed for that protocols may also be essential for getting ready much better cell lysis. We even more evaluated the DNA preparations from all five procedures based on T RFLP evaluation. The Shannon Weiner index was made use of to indicate diversity and complexity, plus the Simpson index was used to measure abundance. Bacteria and archaea have been analyzed separately. The outcomes showed that Protocol E resulted in the highest bacterial diversity and also the highest abundance, fol lowed by Protocols P and EY. Protocols E and EY showed higher archaeal enrichment than that of Protocols P, F and S.
It might protect against CCl4 induced toxicity and oxidative anxiety. The mecha nism of protection involves decreased production of ROS and lipid peroxidation. The agnuside, vitegnoside and fla vonoids existing from the plants are also organic antioxidants. VN extract showed sizeable levels of percent inhibition of DPPH? compared on the standard antioxi dants which had been used as good control. As proven in this table, VN extract was drastically reduce than BHT, galic acid and ascorbic acid at minimal concentrations, three. 125 and six. 25 ug ml offering lowest inhibition at 2. 35% and 2. 58%, respectively. Interestingly, this inhibition was sig nificantly improved from the expanding the concentration from 12. 3 to 25 to 50 ug ml to offer 79. 43% with IC50 13. 31 0. 18 ug L. This inhibition is also closed to BHT 82.
53% with IC50 13. 8 0. 14 ug L and gallic acid 89. 51% with IC50 3. one 0. 08 ug L. This plainly signifies that the VN extract has excellent radical scavenging activity compared towards the pure compounds. Presumably, signifi cantly greater inhibition of DPPH might be attributed towards the presence of multi hydroxyl groups, that’s the ac tive center of anti oxidation selleck like7, 8 dimethyl herbacetin 3 rhamnoside and vitegnoside which showed radical scavenging action 97. 3% and 95. 6% respectively. Even though we usually do not have an exact explanation to the greater maximize of VN no cost radical scavenging action, we could offer some logical arguments. Our published information showed that VN is rich in phenolic compounds and offers a wide variety of antioxidant properties which seems to be straight related to the hydroxyl groups at tached to aromatic rings.
This broad spectrum of antioxi dant formula provides the ideal probable protection towards the zero cost radicals. This attributed to bond dissociation en ergy of every hydroxyl group attached towards the ben zene ring. It can be the power required to break one mole on the bond to offer separated atoms. JNJ26481585 BDE plays a central purpose in figuring out antioxidant efficacy, and it is among the many most critical physical parameters employed for evaluating antioxi dant action in chemical compounds which have been implemented as inhibitors of free radical reactions. Normally, com pounds acquiring lower BDEs are already reported to get much better antioxidant properties. Hence, VN has a wide selection of hydroxyl groups and thus, exerts a wide range of BDEs, forming a synergistic process involving antioxidant and co antioxidant by regeneration of antioxidants by means of the co antioxidant. For example, vitamin E and polyphenols could be concerned while in the synergistic antioxidant method. TOH is consumed from your starting with the oxidation reaction forming TO? radical which com pletely preserved until eventually all co antioxidant is consumed.
Having said that, during the literature there’s disagreement regard ing the prognostic influence of Akt expression. Although the vast majority of scientific studies agree that Akt expression overtly signifies a bad prognosis 19 21, there are plenty of stu dies displaying the opposite impact 22,23. Expressions of PI3K/Akt pathway elements have rarely been investi gated in STSs and you will discover virtually no scientific studies devoted to their prognostic worth 24. Distinctive physiological perform from the Akt household kinases implies that the expression of its isoforms may also have numerous prognostic effect in cancer. The sig nificance of this variation for the survival within the STS sufferers is not nicely investigated and its not clear if the web-site of phosphorylation as well as the pattern of expression can perform prognostic roles.
In prior research, we have now shown the prognostic worth of female steroid hormone receptors in STSs, each alone and while in the coexpression with TGF b and fascin 25,26. Such prognostic affect is not really surprising, considering the fact that both ER and PgR regulate growth and cell differentiation upon ligand dependent and ligand independent activa tion and therefore are in essence growth aspects. Within this context we selleck chemicals desired to examine the correlations concerning female hormone receptors as well as the members of PI3K/Akt sig naling pathway. To our information, these correlations haven’t been described previously. On this examine, we investigate the prognostic impact of all isoforms of Akt, PI3K, PTEN, ER and PgR in 249 non GIST STS individuals. GIST scenarios were excluded from your examine given that sufferers with this subtype of sarcoma receive a specific treatment routine which resulted in drastically considerably better survival.
Resources and solutions Sufferers and clinical samples Primary tumor tissue from anonymized patients diag nosed with non GIST STS at the University Hospital of North Norway 1973 2006 and the Hospitals of Arkhangelsk region, Russia, have been utilized in this retro spective research. In complete, 496 patients had been registered through the hospital knowing it databases. Of these, 247 patients had been excluded thanks to missing clinical data or inadequate materials for histological examination. Consequently, 249 STS patients with complete clinical information and adequate paraffin embedded tissue blocks have been eligible. This report includes observe up information as of September 2009. The median stick to up was 38 months. Formalin fixed and paraffin embedded tumor specimens have been obtained from the archives of the Departments of Pathology at UNN as well as the Arkhangelsk hospitals.
The tumors had been graded in accordance to the French F?d?ration Nationales des Centres de Lutte Con tre le Cancer 27. Microarray development All sarcomas had been histologically reviewed by two educated pathologists along with the most representative parts of viable tumor cells were care thoroughly chosen and marked to the hematoxylin and eosin stained slides and sampled for the tissue micro array blocks.
PLC signaling is very important for PDGF BB induced Akt phosphorylation To verify our acquiring that Ca2 is involved in regula tion of Akt phosphorylation on Ser473, we utilised domin ant damaging PLC?, as well as lower molecular bodyweight inhibitor U73122, which inhibits each PLC? and PLD. Consistent with all the effect of Ca2 chelation, U73122, as well as dnPLC? inhibited Ser473 phosphorylation on Akt, having said that, no impact for the phos phorylation of Thr308 was uncovered. Moreover, U73122 also inhibited S6 phosphorylation, in concurrence together with the ability of this drug to inhibit PLD. To even more investigate the position of PLC? signaling in Akt activation, we utilised PLC?1 null cells. Importantly, these cells have been shown to also have a deficient PLD acti vation. Implementing these cells, we observed a defect in PDGF BB induced Akt phosphorylation on Ser473, but also on Thr308.
This surprising choosing suggests that phosphorylation of Akt on Ser473 is selleck chemical dependent on PLC? action, whereas the phosphoryl ation on Thr308, that’s not impacted by PLC inhibition or Ca2 chelation, necessitates the presence of PLC?one, but not always its exercise. Previously, it’s been shown that inhibition of p38 signaling by SB203580 lowers Akt phosphorylation. This effect was not observed in our experiment. Given that PKC isoforms are activated downstream of PLC?, and it has been reported that mTORC2 regulates the stability and phosphorylation of PKC, we investigated in the event the necessity of Ca2 and PLC? for Akt phosphorylation occurred by activation of PKC. Initial, we confirmed the previously reported reduc tion of PKC levels inside the Rictor null cells.
Upcoming, we downregulated the PKC isoforms which might be dependent on diacylglycerol for their activation, by treating cells with PMA overnight. To monitor the effect of PMA treatment, we investigated phosphorylation of Myristoylated Alanine Wealthy C Kinase, AZ628 a acknowledged PKC substrate. In cells with downre gulated PKC isoforms, we observed a partial reduction inside the ability of PDGF BB to advertise Akt phosphorylation. Steady with our former experiments, we found that S6 phosphorylation was independent of your reduction in Akt phosphorylation. The action of PLC? has become connected to its phos phorylation on Tyr783. To check out in case the absence of Rictor has an effect on PLC? perform, we analyzed the ability of PDGF BB to stimulate PLC? phosphorylation.
Remarkably, we identified that in Rictor null cells the PLC? phosphorylation was abolished and just like what was observed for PKC, the total protein level was somewhat decreased. The mechanism for the diminished PLC? protein level is unclear, but inside the case of PKC it has been demonstrated that mTOR mediated phosphorylation is vital for protein sta bility. To conclude, inhibition of PLC? or Ca2 chelation resulted in decreased PDGF BB induced phosphory lation of Akt on Ser473, but didn’t affect phosphory lation on Thr308.
The total OTUs for all phyla had dropped to 67% of your October 2010 level. From the time period from April 2011 to October 2011, numerous within the bacterial phyla that had a decrease in OTUs through the proceeding period started to recover. Actinobacteria, Firmicutes, and Spirochaetes all had increased numbers of OTUs, and as a percentage of total OTUs they had all surpassed their October 2010 ranges. Proteobacteria was even now probably the most abundant phylum nevertheless it represented only 39% with the complete OTUs in October 2011. The B proteobacterial class had drastically selleck chemical more OTUs as did the purchase Burkholderiales, The quantity of OTUs while in the proteobacterial class decreased drastically, and this difference appears concentrated while in the buy Enterobacteriales, Although the bacterial OTU amounts appeared to get trending upward, by October of 2011 the general abundance of bacteria was nevertheless only 72% in the October 2010 degree.
Las bacterium in HLB impacted citrus handled with antibiotic combinations The dynamic variations of Las bacterial titers from August 2010 to October 2011 on the USHRL farm, Fort Pierce, FL are presented in Figure 1. The results LY2835219 concentration showed that the Las bacterial population fluctuated through the entire year in HLB affected citrus plants with or with no antibiotic deal with ments. The highest Las bacterial titers had been observed in December 2010, as well as the lowest Las bacterial titers have been recorded in April 2011. This variation usually coincided with HLB symptoms from the area. There were no sizeable differ ences amongst the antibiotic therapies along with the water con trol prior to the first applications in August 2010.
Two months after the initial applications, major distinctions existed between the antibiotic treatment options as well as controls. By April 2011, the titers had decreased by over 13 fold during the water manage, 259 fold from the KO handled citrus and 97 fold while in the PS handled citrus. The HybScore of OTU63806, which represented Candidatus Liberibacter from PhyloChip G3, coincided together with the Las bacterial titers detected by qPCR, HybScores av eraged 12,186 one,320 while in the untreated trees when compared with eleven,226 one,458 and 11,037 678 inside the HLB impacted trees taken care of with KO and PS, respectively. HybScores have been the lowest in April 2011 when the HLB bacterial population was also at its lowest level, Bacterial neighborhood framework and diversity The PhyloChip G3 array was implemented to achieve insights to the structural composition and diversity of bacteria in the itrus treated with anti biotic combinations, Of the seven,028 OTUs from our area citrus samples observed about the PhyloChip G3, a complete of 5,599 had been detected in our antibiotic handled discipline samples.
Thus, the initiator methionine is not really the one particular indicated in the database, as well as the protein is 298 amino acids. Surprisingly, there may be no evident Shine Dalgarno sequence adjacent on the initiator methionine we recognized, Phenotype of the S. flexneri gluQ rs mutant To determine the role of GluQ RS in S. flexneri development and virulence, a deletion mutant with the gluQ rs gene was constructed in S. flexneri 2457T. The mutant was com pared for the wild variety by Biolog phenotype MicroArrays, The key distinction observed for your mutant was impaired metabolic process when grown underneath osmotic anxiety situations, The mutant had a longer lag and diminished development in contrast for the wild style inside the presence of increasing concentra tions of potassium chloride, sodium sulfate, sodium formate, sodium benzoate, sodium nitrate and sodium nitrite.
The phenotype was complemented with the gluQ rs gene cloned into an expression vector. No distinctions had been observed during the development or metabolic process of those strains whenever they had been incubated in presence of 1% sodium chloride, which was much like LB, Since expression of dksA is required for S. flexneri virulence, and development of Shigella in the intracellular chk2 inhibitor atmosphere could possibly induce a tension response, we also mea sured invasion and plaque formation through the gluQ rs mutant. Nonetheless, no major distinctions had been mentioned, suggesting that GluQ RS is just not essen tial for invasion or intracellular development of S. flexneri. Discussion Conserved dksA gluQ rs genomic organization in gammaproteobacteria GluQ RS, a paralog of GluRS synthetase, is involved with the formation of GluQ, the nucleoside located with the wob ble position of tRNAAsp in bacteria.
The protein is present in Firmicutes, Actinobacteria, Cyanobacteria, Alphapro Oridonin teobacteria, Betaproteobacteria, Gammaproteobacteria and Deltaproteobacteria, From the phylogenetic evaluation we distinguished the 3 subgroups described previously based mostly on the High motif which is existing inside the class I aminoacyl tRNA synthetases, As was described previously, all GluQ RS enzymes are charac terized by the replacement of the threonine in GluRS enzymes, which is associated with the recognition with the amino acid as well as the terminal adenosine within the tRNAGlu by isoleu cine, leucine or valine at that place, This substitution can be conserved in all enzymes analyzed right here, as well as individuals from your Firmicutes group.
The gluQ rs gene is broadly distributed within the bacterial do most important. even so, its genome organization is variable. We observed that only in members of your gammaproteobacteria, namely Aeromonadales, Alteromonadales, Pseudomonadaceae, Enterobacteriaceae and Vibrionaceae, the gluQ rs gene is found without delay downstream of your dksA gene, A much more thorough evaluation shows that even within this genomic organization there are actually differences.