ArticleMethods in molecular biology (Clifton, N.J.)2013
Isolation of bacterial type IV machine subassemblies.
Article in Methods in molecular biology (Clifton, N.J.), 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Article
- Enhanced Biofilm Formation and Membrane Vesicle Release byFrontiers in microbiology · 2018Article
- The Mosaic Type IV Secretion Systems.EcoSal Plus · 2016Review
- Article
- The Agrobacterium Ti Plasmids.Microbiology spectrum · 2014Review
- Mechanism and structure of the bacterial type IV secretion systems.Biochimica et biophysica acta · 2014Review
- Echoes of a distant past: The cag pathogenicity island of Helicobacter pylori.Cold Spring Harbor perspectives in medicine · 2013Review
- A putative transmembrane leucine zipper of agrobacterium VirB10 is essential for t-pilus biogenesis but not type IV secretion.Journal of bacteriology · 2013Article
- DNA substrate-induced activation of the Agrobacterium VirB/VirD4 type IV secretion system.Journal of bacteriology · 2013Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
The bacterial type IV secretion systems (T4SSs) deliver DNA and protein substrates to bacterial and eukaryotic target cells generally by a mechanism requiring direct contact between donor and target cells. Recent advances in defining the architectures of T4SSs have been made through isolation of machine subassemblies for further biochemical and ultrastructural analysis. Here, we describe a protocol for isolation and characterization of VirB protein complexes from the paradigmatic VirB/VirD4 T4SS of Agrobacterium tumefaciens. This protocol can be adapted for isolation of T4SS subassemblies from other gram-negative bacteria as well as gram-positive bacteria. The biological importance of isolated T4SS subcomplexes can be assessed by assaying for copurification of trapped or cross-linked substrates. This can be achieved with a modified form of the chromatin immunoprecipitation (ChIP) assay termed transfer DNA immunoprecipitation (TrIP). Here, a TrIP protocol is described for recovery of formaldehyde-cross-linked DNA substrate-channel subunit complexes from cells employing T4SSs for conjugative DNA transfer.
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