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SLICER:A Seamless Gene Deletion Method for Deinococcus radiodurans

BioDesign Research 2023年第1期5卷 62-71页

作者： Stephanie L.Brumwell Katherine D.Van Belois Daniel p.Nucifora bogumil J.Karas Department of Biochemistry Schulich School of Medicine and DentistryThe University of WesternOntarioLondonON N6A 5C1Canada. Department of Biology The University of Western OntarioLondonONN6A 5B7Canada.

Deinococcus radiodurans’high resistance to various stressors combined with its ability to utilize sustainable carbon sources makes it an attractive bacterial chassis for synthetic biology and industrial ***,to fully harness the capabilities of this microbe,further strain engineering and tool development are *** for creating seamless genome modifications are an essential part of the microbial genetic toolkit to enable strain ***,we report the development of the SLICER method,which can be used to create seamless gene deletions in *** process involves(a)integration of a seamless deletion cassette replacing a target gene,(b)introduction of the pSLICER plasmid to mediate cassette excision by I-SceI endonuclease cleavage and homologous recombination,and(c)curing of the helper *** demonstrate the utility of SLICER for creating multiple gene deletions in *** by sequentially targeting 5 putative restriction-modification system genes,recycling the same selective and screening markers for each subsequent *** we observed no significant increase in transformation efficiency for most of the knockout strains,we demonstrated SLICER as a promising method to create a fully restriction-minus strain to expand the synthetic biology applications of ***,including its potential as an in vivo DNA assembly platform.

关键词： radiodurans utilize attractive

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photometric observations of three high mass X-ray binaries and a search for variations induced by orbital motion

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Research in Astronomy and Astrophysics 2011年第8期11卷 947-964页

作者： Gordon E. Sarty bogumil p.lecki Daniel E. Reichart Kevin M. Ivarsen Joshua B. Haislip Melissa C. Nysewander Aaron p. LaCluyze Helen M. Johnston Robert R. Shobbrook Lászl L. Kiss Kinwah Wu Departments of psychology and physics University of Saskatchewan9 Campus DriveSaskatoonSaskatchewan S7N 5A5Canada Warsaw University Observatory Al. Ujazdowskie 400-478 WarsawPoland Universidad de Concepcion Departamento de AstronomaCasilla 160-CConcepcionChile Department of physics and Astronomy University of North Carolina-Chapel HillCB 3255Phillips HallChapel HillNC 27599-3255USA School of physics A28 University of SydneyNew South Wales 2006Australia Konkoly Observatory H-1525 BudapestP.O. Box 67Hungary Mullard Space Science Laboratory University College LondonHolmbury St. MaryDorkingSurrey RH5 6NTUnited Kingdom

We searched for long period variation in V-band, It-band and RXTE X-ray light curves of the High Mass X-ray Binaries （HMXBs） LS 1698 / RX J1037.5-5647, HD 110432 / 1H 1249-637 and HD 161103 / RX J1744.7-2713 in an attempt to discover orbitally induced variation. Data were obtained primarily from the ASAS database and were supplemented by shorter term observations made with the 24- and 40-inch ANU telescopes and one of the robotic pROMpT telescopes. Fourier periodograms suggested the existence of long period variation in the V-band light curves of all three HMXBs, however folding the data at those periods did not reveal convincing periodic variation. At this point we cannot rule out the existence of long term V-band variation for these three sources and hints of longer term variation may be seen in the higher precision pROMpT data. Long term V-band observations, on the order of several years, taken at a frequency of at least once per week and with a precision of 0.01 mag, therefore still have a chance of revealing long term variation in these three HMXBs.

关键词： accretion -- stars: Be stars -- stars: neutron stars -- stars: individual LS 1698 HD 110432 HD 161103 -- X-rays: binaries

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Superior Conjugative plasmids Delivered by Bacteria to Diverse Fungi

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BioDesign Research 2022年第1期2022卷 151-165页

作者： Ryan R.Cochrane Arina Shrestha Mariana M.Severo de Almeida Michelle Agyare-Tabbi Stephanie L.Brumwell Samir Hamadache Jordyn S.Meaney Daniel p.Nucifora Henry Heng Say Jehoshua Sharma Maximillian p.M.Soltysiak Cheryl Tong Katherine Van Belois Emma J.L.Walker Marc-AndréLachance Gregory B.Gloor David R.Edgell Rebecca S.Shapiro bogumil J.Karas Department of Biochemistry Schulich School of Medicine and DentistryThe University of Western OntarioLondonONCanada N6A 5C1 Department of Molecular and Cellular Biology University of GuelphGuelphONCanada N1G 2W1 Department of Biology The University of Western OntarioLondonOntarioCanada N6A 5B7

Fungi are nature’s recyclers,allowing for ecological nutrient cycling and,in turn,the continuation of life on *** fungi inhabit the human microbiome where they can provide health benefits,while others are opportunistic pathogens that can cause ***,members of the fungal kingdom,have been domesticated by humans for the production of beer,bread,and,recently,medicine and ***,the great untapped potential exists within the diverse fungal ***,many yeasts are intractable,preventing their use in biotechnology or in the development of novel treatments for pathogenic ***,as a first step for the domestication of new fungi,an efficient DNA delivery method needs to be ***,we report the creation of superior conjugative plasmids and demonstrate their transfer via conjugation from bacteria to 7 diverse yeast species including the emerging pathogen Candida *** create our superior plasmids,derivatives of the 57 kb conjugative plasmid pTA-Mob 2.0 were built using designed gene deletions and insertions,as well as some unintentional ***,a cluster mutation in the promoter of the conjugative gene traJ had the most significant effect on improving conjugation to *** addition,we created Golden Gate assembly-compatible plasmid derivatives that allow for the generation of custom plasmids to enable the rapid insertion of designer genetic ***,we demonstrated that designer conjugative plasmids harboring engineered restriction endonucleases can be used as a novel antifungal agent,with important applications for the development of next-generation antifungal therapeutics.

关键词： insertion continuation harbor

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