Synthetic Core Promoters as Universal Parts for Fine-Tuning Expression in Different Yeast Species

Synthetic biology and metabolic engineering experiments frequently require the fine-tuning of gene expression to balance and optimize protein levels of regulators or metabolic enzymes. A key concept of synthetic biology is the development of modular parts that can be used in different contexts. Here, we have applied a computational multifactor design approach to generate <i>de novo</i> synthetic core promoters and 5′ untranslated regions (UTRs) for yeast cells. In contrast to upstream <i>cis</i>-regulatory modules (CRMs), core promoters are typically not subject to specific regulation, making them ideal engineering targets for gene expression fine-tuning. 112 synthetic core promoter sequences were designed on the basis of the sequence/function relationship of natural core promoters, nucleosome occupancy and the presence of short motifs. The synthetic core promoters were fused to the <i>Pichia pastoris AOX1</i> CRM, and the resulting activity spanned more than a 200-fold range (0.3% to 70.6% of the wild type <i>AOX1</i> level). The top-ten synthetic core promoters with highest activity were fused to six additional CRMs (three in <i>P. pastoris</i> and three in <i>Saccharomyces cerevisiae</i>). Inducible CRM constructs showed significantly higher activity than constitutive CRMs, reaching up to 176% of natural core promoters. Comparing the activity of the same synthetic core promoters fused to different CRMs revealed high correlations only for CRMs within the same organism. These data suggest that modularity is maintained to some extent but only within the same organism. Due to the conserved role of eukaryotic core promoters, this rational design concept may be transferred to other organisms as a generic engineering tool

Sport Aspects in Basketball Training of Children and Youth in Lithuania and Czech Republic

Title: Sport aspects in basketball training of children and youth in Lithuania and Czech republic Work aims: To find differences or common trakte in trainnig children and youth and to gain information needed to raise the duality of basketball as one. The search for welfare mistakes in development of Czech children. To get close to the Lithuanian training mock u pand aspects of physical education of players. Method: Method of questioning by form of applications (questions with available answers). Personal views and thoughts acquired from own experience in Czech schools and Czech teams in comparison and experinence in Lithuania on as exchange program in 2007. Results: With this diploma thesis we would like to contribute to work improvement with young people and try to bring my ideas closer to others slony with training philosphy and physical education. Based on basic guidelines to show or kontrast views of Czech and Lithuianian coaches who worked or work with young peopla and to find out where the differences if theraq are. To make Czech coaches understand Lithuanian view of basketball training. Keywords: basketball, children and youth, trainni

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-8

Ns. Full symbols represent parameter estimation results and open symbols the model validation results. Dashed lines are the 95% confidence limits for the parameter estimation and model validation results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-3

Eeding conditions. Full symbols represent the experimental data and full lines the modelling results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-0

Ns. Full symbols represent parameter estimation results and open symbols the model validation results. Dashed lines are the 95% confidence limits for the parameter estimation and model validation results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-4

Tions. Full symbols represent the experimental data and full lines the modelling results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-5

E feeding conditions. Full symbols represent the experimental data and full lines the modelling results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-1

Nditions. Full symbols represent parameter estimation results and open symbols the model validation results. Dashed lines are the 95% confidence limits for the parameter estimation and model validation results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-7

Tent of 24% (C-mol basis).<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures-10

Ions. Full symbols represent the experimental data and full lines the modelling results.<p><b>Copyright information:</b></p><p>Taken from "Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures"</p><p>http://www.biomedcentral.com/1752-0509/2/59</p><p>BMC Systems Biology 2008;2():59-59.</p><p>Published online 8 Jul 2008</p><p>PMCID:PMC2483998.</p><p></p