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Some of Our Methods and Techniques Employed

While standard biochemistry and molecular biological techniques suffice for much of our work, several of our studies require sophisticated state-of-the-art equipment that is made available to us at the California NanoSystems Institute (CNSI) here at UCLA or in the laboratories of our collaborators.

Physical/chemical: fluorescence correlation spectroscopy (FCS); high resolution cryo-electron microscopy (cryo-EM); magnetic and optical tweezers (Professors James Bowie (UCLA) and Doug Smith (UCSD)) maleimide and “click” chemistries for conjugating ligands to capsid protein; electrophoretic, sedimentation, and chromatographic separations and analyses of fluorescently-labeled RNA, protein, and RNA-protein complexes; labeling of RNA ends, and of capsid proteins, by <2nm gold particles; statistical-mechanical modeling.

Biological: agrobacterial transformation of plants for high-level expression of wildtype and mutant CCMV capsid protein; genetic engineering of RNA-virus-derived replicons from mammalian, insect, and plant viruses; transfection and infection of cultured cells for assays of RNA replication and protein expression levels; in vitro transcription and genetic engineering of viral genes and their protein products; cell-free translation of RNA, of virus-like particles, and of viruses.