Aims
Using substrate‐induced gene‐expression (SIGEX) screening on subseafloor sediment samples from the Nankai Trough, Japan, we identified gene fragments showing an induction response to metal ions.
...Methods and Results
Environmental DNA libraries in Escherichia coli host cells were tested by the addition of metal ions (Ni2+, Co2+, Ga3+ or Mo6+), followed by cell sorting of clones exhibiting green fluorescence upon co‐expression of green fluorescence protein downstream of the inserted gene fragments. One clone displayed Ni2+‐specific induction, three clones displayed Ga3+‐specific induction and three clones displayed an induction response to multiple metal ions. DNA sequence analysis showed that a variety of genes showed induction responses in the screened clones.
Conclusions
Using the SIGEX approach, we retrieved gene fragments with no previously identified response to metal ions that exhibited metal‐ion‐induced expression. This method has the potential to promote exploration of gene function through gene‐induction response.
Significance and Impact of the Study
We successfully linked gene‐induction response with sequence information for gene fragments of previously unknown function. The SIGEX‐based approach exhibited the potential to identify genetic function in unknown gene pools from the deep subseafloor biosphere, as well as novel genetic components for future biotechnological applications.
As arrangements, the fundamental solutions of anisotropic convective diffusion equations of transient incompressible viscous fluid flow and boundary elements analysis of the diffusion equation are ...presented. Secondly, by considering that convective diffusion equations and Navier–Stokes equations are mathematical formulations of mass and momentum conservation law respectively, and that consequently, both physical contents and equation styles are analogous, boundary integral formulations for Navier–Stokes equations are proposed on the basis of formulation of diffusion equations.
The present study investigates the human leucocyte antigen (HLA) allele and haplotype frequencies in Japanese population. We carried out the frequency analysis in 5824 families living across Japanese ...archipelago. The studied population has mainly been typed for the purpose of transplant, especially the hematopoietic stem cell transplantation (HSCT). We determined HLA class I (A, B, and C) and HLA class II (DRB1) using Luminex technology. The haplotypes were directly counted by segregation. A total of 44 HLA‐A, 29 HLA‐C, 75 HLA‐B, and 42 HLA‐DRB1 alleles were identified. In the HLA haplotypes of A‐C‐B‐DRB1 and C‐B, the pattern of linkage disequilibrium peculiar to Japanese population has been confirmed. Moreover, the haplotype frequencies based on family study was compared with the frequencies estimated by maximum likelihood estimation (MLE), and the equivalent results were obtained. The allele and haplotype frequencies obtained in this study could be useful for anthropology, transplantation therapy, and disease association studies.
Model-driven development Mellor, Stephen J; Clark, Anthony N; Futagami, Takao
IEEE software,
09/2003, Volume:
20, Issue:
5
Journal Article
Peer reviewed
Open access
Model driven development is simply the notion that people can construct a model of a system that they can then transform into the real thing. In this way, models become assets instead of ...expenses--quite the business proposition.
The General Well-Being Schedule (GWBS) is a widely used, self-administered questionnaire to measure subjective well-being. It contains 18 items and was originally hypothesized to have six domains; ...however, the following studies suggest other factorial models. We devised the schedule's Japanese edition following the standard procedure and tested its psychometric properties considering Japanese cultural characteristics. Subjects were 1224 middle-aged participants undergoing health checkups in Adachi, Tokyo. The reliability of the original version was high: alpha coefficient, 0.90; test-retest Pearson's correlation coefficient, 0.81; and intra-class correlation coefficient, 0.85. Confirmatory factor analyses supported the structural validity of the 6-factor, the following 3-factor and 4-factor models (goodness of fit index: 0.932, 0.946 and 0.937, respectively). A new 3-factor model developed after the exclusion of item 11, which had no correlation with the other variables, was proved to have a higher fitness index (0.958) than the other models. This 17-item version showed adequate reliability (α coefficient, 0.91; test-retest Pearson's correlation coefficient, 0.81; and intra-class correlation coefficient, 0.85) and concurrent validity compared with the General Health Questionnaire, State-Trait Anxiety Inventory, Center for Epidemiologic Studies Depression Scale, Self-rating Depression Scale and Profile Of Mood State (correlation coefficients: -0.75 to -0.53). The median time required to complete the schedule was five minutes. Completing this instrument was easy for 80% of the subjects. These findings attest to the adequacy of the 17-item Japanese GWBS for the comprehensive assessment of individual inner state.
In our previous studies, we demonstrated that chimeric molecules of the CMP-sialic acid (CMP-Sia) transporter (CST) and the UDP-galactose (Gal) transporter (UGT) in which the seventh transmembrane ...helix-containing segment was derived from the CST could transport both CMP-Sia and UDP-Gal and that the CST-derived seventh transmembrane helix segment was sufficient for the chimera to recognize CMP-Sia in the otherwise UGT context. In this study, we continued to more precisely define the submolecular region that is necessary for CMP-Sia recognition, and we demonstrated that the N-terminal half of the seventh transmembrane helix of CST is essential for the CMP-Sia transport mediated by the chimeric transporters. We further showed that Tyr214Gly and Ser216Phe mutations of a chimeric transporter that was capable of transporting both CMP-Sia and UDP-Gal led to the selective loss of CMP-Sia transport activity without affecting UDP-Gal transport activity. Conversely, when a residue in a chimeric transporter that was active for UDP-Gal transport but not CMP-Sia transport was replaced by Tyr, so that Tyr occupied the same position as in the CMP-Sia transporter, the resulting mutant chimera acquired the ability to transport CMP-Sia. These results demonstrated that Tyr214 and Ser216, located in the seventh transmembrane helix of the human CST, are critically important for the recognition of CMP-Sia as a transport substrate. Identification of determinants critical for the discrimination between relevant and irrelevant substrates will advance our understanding of the mechanisms of substrate recognition by nucleotide sugar transporters.
PDF
