Genetics and Cell Morphology Analyses of the Actinomyces oris srtA Mutant.
Sortase is a cysteine-transpeptidase that anchors LPXTG-containing proteins on the Gram-positive bacterial cell wall. Previously, sortase was considered to be an important factor for bacterial pathogenesis and fitness, but not cell growth. However, the Actinomyces oris sortase is essential for cell viability, due to its coupling to a glycosylation pathway. In this chapter, we describe the methods to generate conditional srtA deletion mutants and identify srtA suppressors by Tn5 transposon mutagenesis. We also provide procedures for analyzing cell morphology of this mutant by thin-section electron microscopy. These techniques can be applied for analyses of other essential genes in A. oris.
['Wu|Chenggang|C|', 'Reardon-Robinson|Melissa Elizabeth|ME|', 'Ton-That|Hung|H|']
D000190:Actinomyces D019881:Aminoacyltransferases D001426:Bacterial Proteins D003546:Cysteine Endopeptidases D004251:DNA Transposable Elements D017353:Gene Deletion D020043:Genes, Essential D006360:Heat-Shock Proteins
Actinomyces oris Sortase Glycosylation Tn5 transposon mutagenesis Electron microscopy Allelic exchange