Molecular Biology


    Protocols in Current Issue
    Overrepresentation Analyses of Differentially Expressed Genes in the Smut Fungus Ustilago bromivora during Saprophytic and in planta Growth
    Authors:  Angelika Czedik-Eysenberg, Franziska Rabe, Heinz Ekker, Carmen Czepe and Armin Djamei, date: 08/05/2017, view: 331, Q&A: 0
    [Abstract] We have established the Ustilago bromivoraBrachypodium spp. interaction as a new model pathosystem for biotrophic fungal plant infections of the head smut type (Rabe et al., 2016). In this protocol, the methodology used for comparing gene expression between saprophytic and in planta growth of the fungus is ...
    Advanced Design of Minimalistic Dumbbell-shaped Gene Expression Vectors
    Authors:  Xiaoou Jiang and Volker Patzel, date: 08/05/2017, view: 298, Q&A: 0
    [Abstract] Minimal DNA vectors exclusively comprising therapeutically relevant sequences hold great promise for the development of novel therapeutic regimen. Dumbbell-shaped vectors represent non-viral non-integrating DNA minimal vectors which have entered an advanced stage of clinical development (Hardee et al., 2017). Spliceable introns and DNA ...
    Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters
    Authors:  Sanchari Bhattacharyya, Shimon Bershtein and Eugene I Shakhnovich, date: 07/20/2017, view: 345, Q&A: 0
    [Abstract] This protocol is used to assay the effect of protein over-expression on fitness of E. coli. It is based on a plasmid expression of a protein of interest from an arabinose-regulated pBAD promoter followed by the measurement of the intracellular protein abundance by Western blot along with the measurement of growth parameters of E. coli ...
    GUS Staining of Guard Cells to Identify Localised Guard Cell Gene Expression
    Authors:  Zhao Liu, Wei Wang, Chun-Guang Zhang, Jun-Feng Zhao and Yu-Ling Chen , date: 07/20/2017, view: 794, Q&A: 0
    [Abstract] Determination of a gene expression in guard cells is essential for studying stomatal movements. GUS staining is one means of detecting the localization of a gene expression in guard cells. If a gene is specially expressed in guard cells, the whole cotyledons or rosette leaf can be used for GUS staining. However, if a gene is expressed in both ...
    A Method to Convert mRNA into a Guide RNA (gRNA) Library without Requiring Previous Bioinformatics Knowledge of the Organism
    Author:  Hiroshi Arakawa, date: 05/20/2017, view: 877, Q&A: 0
    [Abstract] While the diversity of species represents a diversity of special biological abilities, many of the genes that encode those special abilities in a variety of species are untouched, leaving an untapped gold mine of genetic information; however, despite current advances in genome bioinformatics, annotation of that genetic information is incomplete in ...
    Fluorescence in situ Localization of Gene Expression Using a lacZ Reporter in the Heterocyst-forming Cyanobacterium Anabaena variabilis
    Authors:  Brenda S. Pratte and Teresa Thiel, date: 01/05/2017, view: 1264, Q&A: 0
    [Abstract] One of the most successful fluorescent proteins, used as a reporter of gene expression in many bacterial, plant and animals, is green fluorescent protein and its modified forms, which also function well in cyanobacteria. However, these fluorescent proteins do not allow rapid and economical quantitation of the reporter gene product, as does the ...
    Single-cell Visualization of Chromosome Transcriptional Territories by RNA-paint
    Authors:  Céline Vallot and Claire Rougeulle, date: 09/05/2016, view: 1683, Q&A: 0
    [Abstract] We developed a FISH-based method to directly assess chromosome-wide transcriptional activity, thereby enabling the visualization of the actively transcribed fraction of a chromosome at the single-cell level. We applied this method to probe the activity of X-chromosomes and its instability in the context of human embryonic stem cells and cancer ...
    Genome-Wide siRNA Screen for Anti-Cancer Drug Resistance in Adherent Cell Lines
    Authors:  Elza C. de Bruin, Ming Jiang, Michael Howell and Julian Downward, date: 05/20/2015, view: 4295, Q&A: 0
    [Abstract] The expression of genes is frequently manipulated in cell lines to study their cellular functions. The use of exogenous small Interfering RNAs (siRNAs) is a very efficient technique to temporarily downregulate the expression of genes of interest [reviewed by Hannon and Rossi (2004)]. A genome-wide siRNA library allows the user to study both the ...
    Random DNA Binding Selection Assay (RDSA)
    Authors:  Yafei Wang and Meizhong Luo, date: 04/20/2015, view: 2329, Q&A: 0
    [Abstract] Protein-DNA interaction is a very important cellular process, by which regulation of DNA biological function, usually gene expression, is exerted. The method of random DNA binding selection assay (RDSA) can be used to identify DNA elements bound by proteins with DNA-binding activities. This method is based on the enrichment of the target DNA ...
    Determination of Luciferase Activity in Arabidopsis seedling
    Authors:  Mohan TC, Gabriel Castrillo and Antonio Leyva, date: 06/20/2014, view: 4107, Q&A: 0
    [Abstract] There are different direct and indirect methods available to study gene expression in plant systems. In this protocol we describe a modified expression assay using transgenic plants expressing the luciferase gene under the control of phosphate transporter PHT1;1 promoter. This assay was originally optimized for analyzing the repression of PHT1;1 ...