3 articles published in JoVE
Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance Chayanika Biswas*1, Sharon C-A. Chen*1,2,3, Catriona Halliday1,2, Elena Martinez1, Rebecca J. Rockett1, Qinning Wang1, Verlaine J. Timms1, Rajat Dhakal1, Rosemarie Sadsad1, Karina J. Kennedy4, Geoffrey Playford4,5, Deborah J. Marriott6, Monica A. Slavin7, Tania C. Sorrell1,3, Vitali Sintchenko1,2,3 1Centre for Infectious Diseases and Microbiology-Public Health, Westmead Hospital, 2Centre for Infectious Diseases and Microbiology Laboratory Services, ICPMR, 3Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, 4Department of Microbiology and Infectious Diseases, Canberra Hospital and Health Services, Australian National University Medical School, 5Infection Management Services, Australian National University Medical School, 6 This study implemented whole genome sequencing for analysis of mutations in genes conferring antifungal drug resistance in Candida glabrata. C. glabrata isolates resistant to echinocandins, azoles and 5-flucytosine, were sequenced to illustrate the methodology. Susceptibility profiles of the isolates correlated with presence or absence of specific mutation patterns in genes.
A Simple Bioassay for the Evaluation of Vascular Endothelial Growth Factors Steven A. Stacker1, Michael M. Halford1, Sally Roufail1, Carol Caesar1, Marc G. Achen1 1Tumour Angiogenesis Program, Peter MacCallum Cancer Centre We describe a simple cell-based bioassay for detecting, quantifying and monitoring the activity of members of the vascular endothelial growth factor family of ligands. The assay uses chimeric receptors expressed in a factor-dependent cell line to provide a semi-quantitative or quantitative assessment of receptor binding and cross-linking by the ligand.
A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl Magdalena Hagn1, Vivien R. Sutton1, Joseph A. Trapani1 1Cancer Immunology Program, Peter MacCallum Cancer Centre We describe a simple, quantitative colorimetric assay that specifically measures the proteolytic activity of human, mouse or rat Granzyme B (GzmB). This protocol can be easily adapted for determining protease activity of other granule serine proteases by the hydrolysis of other synthetic peptide substrates with an appropriate recognition sequence.