Purification of recombinant human and Drosophila septin hexamers for TIRF assays of actin-septin filament assembly
Septins are guanine nucleotide-binding proteins that are conserved from fungi to humans. Septins assemble into heterooligomeric complexes and higher-order structures with key roles in various cellular functions including cell migration and division. The mechanisms by which septins assemble and interact with other cytoskeletal elements like actin remain elusive. A powerful approach to address this question is by cell-free reconstitution of pu- rified cytoskeletal proteins combined with fluorescence microscopy. Here, we describe procedures for the purification of recombinant Drosophila and human septin hexamers from Escherichia coli and reconstitution of actin-septin coassembly. These procedures can be used to compare assembly of Drosophila and human septins and their coassembly with the actin cytoskeleton by total internal reflection fluorescence microscopy.
Code (0)
등록된 구현이 없습니다.
Similar Papers 제목 키워드 기반
Visualizing septins in early Drosophila embryos
Functional studies in Drosophila have been key for establishing a role for the septin family of proteins in animal cell division and thus extending for the first time observations from the budding yeast to animal cells. …
Design and expression of TolC as a recombinant protein vaccine against Shigella flexneri and evaluation of immunogenic response in mice
Background: Shigella is one of the major cause of dysenteric diarrhea, which is known shigelosis. Shigelosis causes 160,000 deaths annually of diarrheal disease in a global scale especially children less than 5 years old…
Simplifying recombinant protein production: Combining Golden Gate cloning with a standardized protein purification scheme
Recombinant protein production is pivotal in molecular biology, enabling profound insights into cellular processes through biophysical, biochemical, and structural analyses of the purified samples. The demand for substan…
Computer-Aided Automated Detection of Gene-Controlled Social Actions of Drosophila
Gene expression of social actions in Drosophilae has been attracting wide interest from biologists, medical scientists and psychologists. Gene-edited Drosophilae have been used as a test platform for experimental investi…
Action DetectionClassificationDimensionality ReductionGeneral Classification+1Recombinant transfer in the basic genome of E. coli
An approximation to the ~4 Mbp basic genome shared by 32 strains of E. coli representing six evolutionary groups has been derived and analyzed computationally. A multiple-alignment of the 32 complete genome sequences was…