HisB as novel selection marker for gene targeting approaches in Aspergillus niger

Background For Aspergillus niger, a broad set of auxotrophic and dominant resistance markers is available. However, only few offer targeted modification of a gene of interest into or at a genomic locus of choice, which hampers functional genomics studies. We thus aimed to extend the available set by generating a histidine auxotrophic strain with a characterized hisB locus for targeted gene integration and deletion in A. niger. Results A histidine-auxotrophic strain was established via disruption of the A. niger hisB gene by using the counterselectable pyrG marker. After curing, a hisB - , pyrG - strain was obtained, which served as recipient strain for further studies. We show here that both hisB orthologs from A. nidulans and A. niger can be used to reestablish histidine prototrophy in this recipient strain. Whereas the hisB gene from A. nidulans was suitable for efficient gene targeting at different loci in A. niger, the hisB gene from A. niger allowed efficient integration of a Tet-on driven luciferase reporter construct at the endogenous non-functional hisB locus. Subsequent analysis of the luciferase activity revealed that the hisB locus is tight under non-inducing conditions and allows even higher luciferase expression levels compared to the pyrG integration locus. Conclusion Taken together, we provide here an alternative selection marker for A. niger, hisB, which allows efficient homologous integration rates as well as high expression levels which compare favorably to the well-established pyrG selection marker.EC/FP7/303864/EU/Bridging the world of fungi and dementia/PROFITSTU Berlin, Open-Access-Mittel - 201

Teaching on Chinese Writing in Binus University

Combination of practice teaching Chinese as a foreign language in BINUS University, this article takes the Chinese writing teaching as an example, and analyses “as the student core, as the fun concept” writing modes. Firstly, serious explain: increasing the vitality of classroom teaching, using multimedia methods to make students get interested in; Secondly, article practice: making the written expression be the basis and revealing true feelings as commander, constantly pursue the aesthetic article; Thirdly, feedback and communion: using Facebook, Binusmaya, Binusblog, Chinese newspapers and other channels of interaction to communicate between teachers and students. Through the three steps, we could select the theme which students loved, create a relaxed atmosphere in the writing class, and then the students will be pleasure of writing gradually

Photoannealing of Microtissues Creates High-Density Capillary Network Containing Living Matter in a Volumetric-Independent Manner

The vascular tree is crucial for the survival and function of large living tissues. Despite breakthroughs in 3D bioprinting to endow engineered tissues with large blood vessels, there is currently no approach to engineer high-density capillary networks into living tissues in a scalable manner. Here, photoannealing of living microtissue (PALM) is presented as a scalable strategy to engineer capillary-rich tissues. Specifically, in-air microfluidics is used to produce living microtissues composed of cell-laden microgels in ultrahigh throughput, which can be photoannealed into a monolithic living matter. Annealed microtissues inherently give rise to an open and interconnected pore network within the resulting living matter. Interestingly, utilizing soft microgels enables microgel deformation, which leads to the uniform formation of capillary-sized pores. Importantly, the ultrahigh throughput nature underlying the microtissue formation uniquely facilitates scalable production of living tissues of clinically relevant sizes (&gt;1 cm3) with an integrated high-density capillary network. In short, PALM generates monolithic, microporous, modular tissues that meet the previously unsolved need for large engineered tissues containing high-density vascular networks, which is anticipated to advance the fields of engineered organs, regenerative medicine, and drug screening.</p

Additional file 5: Figure S5. of HisB as novel selection marker for gene targeting approaches in Aspergillus niger

Growth comparison of olvA - and olvA + mutants with AnidhisB or AopyrG background on CM plates. (TIFF 7346 kb