Protocols in Current Issue
    Quantitative Transformation Efficiency Assay for Bacillus subtilis
    Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 1225, Q&A: 0
    [Abstract] Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...
    Induction of Natural Competence in Genetically-modified Lactococcus lactis
    Authors:  Joyce Mulder, Michiel Wels, Oscar P. Kuipers, Michiel Kleerebezem and Peter A. Bron, date: 07/05/2018, view: 1636, Q&A: 0
    [Abstract] Natural competence can be activated in Lactoccocus lactis subsp lactis and cremoris upon overexpression of ComX, a master regulator of bacterial competence. Herein, we demonstrate a method to activate bacterial competence by regulating the expression of the comX gene by using a nisin-inducible promoter in an L. ...
    Conjugation Assay for Testing CRISPR-Cas Anti-plasmid Immunity in Staphylococci
    Authors:  Forrest C. Walker and Asma Hatoum-Aslan, date: 05/05/2017, view: 5447, Q&A: 0
    [Abstract] CRISPR-Cas is a prokaryotic adaptive immune system that prevents uptake of mobile genetic elements such as bacteriophages and plasmids. Plasmid transfer between bacteria is of particular clinical concern due to increasing amounts of antibiotic resistant pathogens found in humans as a result of transfer of resistance plasmids within and between ...
    Polyethylene Glycol-mediated Transformation of Drechmeria coniospora
    Authors:  Le D. He and Jonathan J. Ewbank, date: 03/05/2017, view: 4515, Q&A: 0
    [Abstract] Drechmeria coniospora is a nematophagous fungus and potential biocontrol agent. It belongs to the Ascomycota. It is related to Hirsutella minnesotensis, another nematophagous fungus but, phylogenetically, it is currently closest to the truffle parasite Tolypocladium ophioglossoides. Together with its natural host, ...
    Single-step Marker Switching in Schizosaccharomyces pombe Using a Lithium Acetate Transformation Protocol
    Authors:  Simon David Brown and Alexander Lorenz, date: 12/20/2016, view: 4631, Q&A: 0
    [Abstract] The ability to utilize different selectable markers for tagging or mutating multiple genes in Schizosaccharomyces pombe is hampered by the historical use of only two selectable markers, ura4+ and kanMX6; the latter conferring resistance to the antibiotic G418 (geneticin). More markers have been described ...
    Transformation of Thermus Species by Natural Competence
    Authors:  Alba Blesa and José Berenguer, date: 11/20/2016, view: 4783, Q&A: 0
    [Abstract] Many Thermus species harbour genomes scourged with horizontally transferred signatures. Thermus thermophilus (Tth) has been characterized as naturally competent. The transformation protocol described here is based on the maximum DNA uptake rates registered at mid-exponential phase (OD600 0.3-0.4). Here we ...
    Cell-to-cell DNA Transfer among Thermus Species
    Authors:  Alba Blesa and José Berenguer, date: 11/20/2016, view: 4823, Q&A: 0
    [Abstract] The ability to transfer DNA via direct cell-to-cell contact-dependent process similar to conjugation has been described in Thermus thermophilus (Tth). Here, we detail the mating experiment protocol involving the lateral transfer of thermostable antibiotic resistance markers (i.e., kanamycin: KmR; hygromycin: HygR ...
    Target Gene Inactivation in Cyanobacterium Anabaena sp. PCC 7120
    Authors:  Kangming Chen, Huilan Zhu, Liping Gu, Shengni Tian and Ruanbao Zhou, date: 08/05/2016, view: 5192, Q&A: 0
    [Abstract] Anabaena sp. strain PCC 7120 has long served as a model organism for investigating N2-fixation, photosynthesis, and various plant-type metabolic pathways and biofuel production, as well as cellular differentiation (Xu et al., 2008, Halfmann et al., 2014, Golden and Yoon, 2003). Since more than 30,000 sequenced ...
    Transformation of the Cyanobacterium Leptolyngbya boryana by Electroporation
    Authors:  Ryoma Tsujimoto, Hiroya Kotani, Aoi Nonaka, Yuri Miyahara, Yuto Hiraide and Yuichi Fujita, date: 12/20/2015, view: 6448, Q&A: 0
    [Abstract] Leptolyngbya boryana (L. boryana) (formerly Plectonema boryanum) is a versatile, filamentous cyanobacterium that has the ability to fix nitrogen under microoxic conditions and to grow heterotrophically with glucose in the dark, providing an excellent system to investigate photosynthesis, nitrogen fixation, and their ...
    Genetic Transformation of Candida glabrata by Heat Shock
    Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 5708, Q&A: 0
    [Abstract] Here, we report a method for the transformation of Candida glabrata using a heat shock method. The protocol can be used for transformations in single well or in 96-well scale. It has been employed as an alternative method to the electroporation protocol to construct a genome-scale gene deletion collection in the human fungal pathogen ...

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