In JoVE (1)
Articles by N. Bishara Marzook in JoVE
Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins N. Bishara Marzook*1, Dean J. Procter*1, Helena Lynn1, Yui Yamamoto2, Jacquelyn Horsington3, Timothy P. Newsome1 1School of Molecular Bioscience, University of Sydney, 2Department of Medicine, Center for Vascular Research, 3Asia Pacific Centre for Animal Health, Faculty of Veterinary Science, University of Melbourne A rapid and modular protocol for the generation of recombinant vaccinia viruses expressing fluorescently tagged proteins simultaneously using the method of transient dominant selection is described here.
Other articles by N. Bishara Marzook on PubMed
Mass Spectrometric Characterization of the Campylobacter Jejuni Adherence Factor CadF Reveals Post-translational Processing That Removes Immunogenicity While Retaining Fibronectin Binding Proteomics. Jan, 2010 | Pubmed ID: 19941310 Campylobacter jejuni is a major gastrointestinal pathogen that colonizes host mucosa via interactions with extracellular matrix proteins, such as fibronectin (Fn). Fn-binding is mediated by a 37 kDa outer membrane protein termed Campylobacter adherence Factor (CadF). The outer membrane protein profile of a recent gastrointestinal C. jejuni clinical isolate (JHH1) was analysed using 2-DE and MS. Several spots were identified as products of the cadF gene. These included mass and pI variants of 34 and 30 kDa, as well as 24 kDa (CadF(24)) and 22 kDa (CadF(22)) mass variants. CadF variants were fully characterized by MALDI-TOF MS and MALDI-MS/MS. These data confirmed that CadF forms re-folding variants resulting in spots with lower mass and varying pI that are identical at the amino acid sequence level and are not modified post-translationally. CadF(22) and CadF(24), however, were characterized as N-terminal, membrane-associated polypeptides resulting from cleavage between serine(195) and leucine(196), and glycine(201) and phenylalanine(202), respectively. These variants were more abundant in the virulent (O) isolate of C. jejuni NCTC11168 when compared with the avirulent (genome sequenced) isolate. Hexahistidine fusion constructs of full-length CadF (34 kDa), CadF(24), and the deleted C-terminal OmpA domain (14 kDa; CadF(14)) were created in Escherichia coli. Recombinant CadF variants were probed against patient sera and revealed that only full-length CadF retained reactivity. Binding assays showed that CadF(24) retained Fn-binding capability, while CadF(14) did not bind Fn. These data suggest that the immunogenic epitope of CadF is cleaved to generate smaller Fn-binding polypeptides, which are not recognized by the host humoral response. CadF cleavage therefore may be associated with virulence in C. jejuni.
Proteomic Profiling of Pseudomonas Aeruginosa AES-1R, PAO1 and PA14 Reveals Potential Virulence Determinants Associated with a Transmissible Cystic Fibrosis-associated Strain BMC Microbiology. 2012 | Pubmed ID: 22264352 Pseudomonas aeruginosa is an opportunistic pathogen that is the major cause of morbidity and mortality in patients with cystic fibrosis (CF). While most CF patients are thought to acquire P. aeruginosa from the environment, person-person transmissible strains have been identified in CF clinics worldwide. The molecular basis for transmissibility and colonization of the CF lung remains poorly understood.