Molecular Biology

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    Protocols in Current Issue
    Quantitative Transformation Efficiency Assay for Bacillus subtilis
    Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 1257, 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: 1660, 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. ...
    Transient Gene Expression for the Characteristic Signal Sequences and the Estimation of the Localization of Target Protein in Plant Cell
    [Abstract] We have proposed and tested a method for characterization of the signal sequences and determinations of target protein localization in a plant cell. This method, called the AgI-PrI, implies extraction of protoplasts from plant tissues after agroinfiltration. The suggested approach combines the advantages of two widely used methods for transient ...
    Generation and Selection of Transgenic Olive Plants
    Authors:  Elena Palomo-Ríos, Sergio Cerezo, Jose Ángel Mercado and Fernando Pliego-Alfaro, date: 11/20/2017, view: 3645, Q&A: 0
    [Abstract] Olive (Olea europaea L.) is one of the most important oil crops in the Mediterranean basin. Biotechnological improvement of this species is hampered by the recalcitrant nature of olive tissue to regenerate in vitro. In previous investigations, our group has developed a reliable Agrobacterium-mediated transformation ...
    Generation of Caenorhabditis elegans Transgenic Animals by DNA Microinjection
    Authors:  Matthias Rieckher and Nektarios Tavernarakis, date: 10/05/2017, view: 5595, Q&A: 0
    [Abstract] Microinjection is the most frequently used tool for genetic transformation of the nematode Caenorhabditis elegans, facilitating the transgenic expression of genes, genome editing by the clustered regularly interspersed short palindromic repeats (CRISPR)-Cas9 system, or transcription of dsRNA for RNA intereference (RNAi). Exogenous DNA is ...
    Ensifer-mediated Arabidopsis thaliana Root Transformation (E-ART): A Protocol to Analyse the Factors that Support Ensifer-mediated Transformation (EMT) of Plant Cells
    Authors:  Dheeraj Singh Rathore, Fiona M. Doohan and Ewen Mullins, date: 10/05/2017, view: 4318, Q&A: 0
    [Abstract] Ensifer adhaerens OV14, a soil borne alpha-proteobacteria of the Rhizobiaceae family, fortifies the novel plant transformation technology platform termed ‘Ensifer-mediated transformation’ (EMT). EMT can stably transform both monocot and dicot species, and the host range of EMT is continuously expanding across a diverse range of ...
    Conjugation Assay for Testing CRISPR-Cas Anti-plasmid Immunity in Staphylococci
    Authors:  Forrest C. Walker and Asma Hatoum-Aslan, date: 05/05/2017, view: 5478, 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 ...
    Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration
    Authors:  Kangquan Yin, Ting Han and Yule Liu, date: 04/05/2017, view: 6826, Q&A: 0
    [Abstract] CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species (Wang et al., 2016). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end ...
    Polyethylene Glycol-mediated Transformation of Drechmeria coniospora
    Authors:  Le D. He and Jonathan J. Ewbank, date: 03/05/2017, view: 4533, 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, ...
    Transformation of Thermus Species by Natural Competence
    Authors:  Alba Blesa and José Berenguer, date: 11/20/2016, view: 4802, 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 ...



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