Keywordsalcohol oxidase 1 (AOX1) promoter, Cat8, engineered promoter, ethanol utilization pathway, Pichia pastoris (Komagataella phaffii)

No abstract

In today's education system, students began to be the subject of the learning environment instead of to be the object of it. Strategies that will provide student participation in the courses also have become even more important in today's education. In this study, the usability of "Transformation Wheel" material that students can work actively participating in the subject of transformation geometry was analyzed. Our study was conducted with the aim of analyzing the usability of "Transformation Wheel" material designed in accordance with the acquisitions of 7th grade mathematics course curriculum about Transformation Geometry and determining the teachers and students' opinions about developed material. For this purpose, the research was carried out with 21 7th graders from a middle school and 9 math teachers who were introduced the material in Istanbul province. In the study, action research method was used. The students were presented to the subject of Transformation Geometry with Transformation Wheel material, then were given worksheets and asked to check their answers that are related to the subject from the Transformation Wheel material. With this way, each student was provided to use the material. After the application, each student's views regarding the material were taken in written. The views of the teachers were also received about the material. And most of the teachers found it to be usable and liked it. As a result of the study, in teaching of mathematical concepts or objectifying the learned concepts, the Transformation Wheel material was found to be effective. It was also determined that it had been applied as studentcentered and had gained students analytical thinking and social interaction skills. In this context, the concrete material of Transformation Wheel was inferred to be usable for teaching mathematics.

Analysis of RNA structuromes provides new insights into cellular processes, enabling systems biology and biotechnology researchers to calculate promoter and terminator strengths and to directly observe how differing circuit states impact host gene expression and the burdens imposed by the circuits. Such analysis, however, is crucially dependent on the availability of highly pure, intact RNA isolated from fresh or frozen cell cultures. RNA extraction from the yeast Pichia pastoris requires specific pretreatment steps to ensure the reproducibility of downstream applications, but current methods and extraction kits are generally adapted for the conventional yeast Saccharomyces cerevisiae, which has a different cell wall composition. We therefore set out to compare the efficacy of two different RNA isolation methods when applied to P. pastoris: (i) phenol/chloroform extraction and (ii) silica spin‐column absorption. We compared the yield, integrity, and purity of the resulting isolated RNA from the two methods (using two different types of commercial columns for silica spin‐column absorption) and further optimized them through variations in the pretreatment steps. We also assessed two different methods of cell lysis: enzyme catalytic disruption using lyticase and mechanical disruption using acid‐washed glass‐beads in a TissueLyser. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: RNA isolation with phenol/chloroform extraction: monophasic lysis reagent Alternate Protocol 1: RNA isolation with silica‐spin column absorption: High Pure RNA Isolation Kit (Roche Life Science) Alternate Protocol 2: RNA isolation with silica‐spin column absorption: RNeasy Mini Kit (Qiagen)