Expression Sample Clauses

Expression. Thus, we employed a mutagenesis analysis to determine whether the inhibitory effect of miR- 324-5p is mediated directly by the putative target region in the 3’UTR of Kv4.2. Accordingly, we designed a reporter construct, ffluc-Kv4.2-3’UTR-MUT, bearing a two nucleotide-mutation in the putative target region of 3’UTR of Kv4.2 (Fig. 3-2A). This point mutation would disrupt the predicted base pairing between miR-324-5p and 3’UTR of Kv4.2. Therefore, the inhibitory effect of miR-324-5p should be eliminated. Indeed, overexpression of miR-324-5p had no significant effect on the luciferase activity of ffluc- Kv4.2-3’UTR-MUT (Fig. 3-2B), suggesting that the target region in 3’UTR is a cis- acting inhibitory element. In line with this observation, ffluc-Kv4.2-3’UTR-MUT exhibits higher luciferase activity than ffluc-Kv4.2-3’UTR (33.0%±8.6%), suggesting that the mutated Kv4.2 construct cannot be regulated by endogenous miR-324-5p (Fig. 3- 2C). Overall, these data suggest that the putative target region of xxXXX-324-5p within the Kv4.2 3’UTR is responsible for the inhibitory function of miR-324-5p on the Kv4.2 reporter constructs, corroborating our hypothesis that miR324-5p directly affects Kv4.2 expression. miR-324-5p is localized to neurons Kv4.2 is highly expressed in the brain and is the predominant potassium channel that composes A-type currents in the hippocampus (Xxxx et al., 2006). Given the potential function of miR-324-5p for Kv4.2 expression, we hypothesized that miR-324- 5p might regulate Kv4.2 in vivo in the brain. We therefore assessed the expression and localization of miR-324-5p in neurons. Fluorescence in situ hybridization (FISH) using digoxigenin labeled LNATM probes was performed to investigate the neuronal localization of miR-324-5p in both brain tissues and cultured hippocampal neurons. miR- 324-5p is localized to both cell bodies and dendrites in the hippocampus of the brain (Fig. 3-3A-C). Correspondingly, miR-324-5p shows expression in 14 day in vitro (DIV) cultured hippocampal neurons (Fig. 3-3D). Overall, these data demonstrate that miR- 324-5p is expressed in neurons and support the hypothesis that miR-324-5p regulates Kv4.2 expression in vivo in the brain. miR-324-5p regulates endogenous Kv4.2 protein levels in primary neurons To investigate whether miR-324-5p modulates the expression of endogenous Kv4.2 protein, we overexpressed or inhibited miR-324-5p in high-density cultured neurons. Lentiviruses expressing both GFP and miR-324-5p or a scr...
Expression. Our results suggest that miR-324-5p may regulate Kv4.2 expression and functions during neuronal development and plasticity. My thesis work provides new insights into the mechanisms regulating neuronal excitability. Dysregulation of these mechanisms might be involved in epileptogenesis, e.g. in FXS. Our findings therefore provide a molecular basis for future studies toward a better understanding the pathogenesis of FXS and other forms of epilepsy.
Expression. At this stage the students practiced the grammatical structures .After his experimental study conducted on the use of is EEE (exploration, explanation and expression) method of L2 teaching grammar teaching he concluded that students preferred to learn grammar through EEE method as compared to only form based or meaning based approaches because they found it to be more effective in learning L2 grammar.
Expression. Students have the right of expression as long as they do not interrupt the educational process. This includes the right to express personal opinions in student publications as long as the statements are not libelous, profane, obscene and do not violate editorial policies governing student publications. Students shall not be required to participate in educational experiences that violate their religious or patriotic convictions, but must request and complete alternative educational experiences. The Academy reserves the right to set reasonable rules regarding freedom of expression. Speech As intellectual beings, students have a right to search vigorously for truth by examining opposing ideas and to espouse and express their views in an orderly manner. A student must be concerned about the effect that his/her spoken word or symbolic expression will have on the personal reputation of others and the reputation of the school; students have the right to have their own personal reputation protected accordingly.
Expression. The terms and expressions referred to in this agreement shall have the same meaning with those used in the Loan Agreement.
Expression. Each of the hybrid transcription units described above will be cloned in the shuttle vector yepl3. Each of the four plasmids will then be transformed into an appropriate leu2 recipient yeast strain. Yeast cultures grown in 4 percent glucose will be assayed for their content of insulin, proinsulin, or related polypeptides using column chromatography, RIA, and gel electrophoresis (western blot) assays. The objective of the research in phase 1 will be to achieve at least a minimal level (0.01 to 0.1 percent of total soluble protein) as intact intracellular insulin/proinsulin, or as a hybridprotein from which intact insulin/proinsulin can easily be obtained by enzymatic or chemical conversion. [*] designates portions of this document that have been omitted pursuant to a request for confidential treatment filed separately with the Commission. The objective of phase II is to construct a genetically stable yeast strain that accumulates intact human insulin/proinsulin or a hybrid protein from which intact insulin/proinsulin can easily be obtained by enzymatic or chemical conversion at [ * ] molecules per 1 at [ * ] cellprotein per 1. By combination of the following approaches this goal will be reached. - Variation in the plasmid vector, particularly as regards its copy number, replication control, segregation mechanism. - Chromosomal integration of the hybrid transcription unit at multiple sites. - Regulatory modulation of the expression of tpi/or other yeast promoter -- both by mutation and by cloning and over-production of gene products required for transcription. - Exploration of the effect of unfavorable codon usage in the proinsulin coding sequence upon expression of proinsulin in yeast. The remedy for this could be to construct a wholly or partially synthetic proinsulin gene with codons which are optimally used in yeast. - Explore the possibilities of using as host organism, other eukaryotic microorganism, that is acceptable for large scale production for NOVO. - Application of general principles for optimizing gene expression in yeast, e.g.: chloramphenical acetyl transferase -- attached in the context of many different sequences at crucial points in the translation unit. Phase III --------- The specific approaches to be followed in optimizing insulin secretion from yeast will depend strongly upon the relative efficacy of various leader peptide constructions used in the phase 1 experiments. In order to increase the level of insulin secretion from yeast, gen...