In-class discussion: Purification: Virus isolation: Characterization: Quantitative methods 2. eclipse phase- production of NA; no complete
particles (3. latent period- general term up to actual
burst) 4. assembly- components packaged +/- "add ons"
(=maturation) 5. burst- release of virons from cell, often violent,
or not Time: bacteriophage- from several seconds to few minutes
to end of latent period [For examples, browse Wagner, ch 2 & 6, and CRC,
ch 3 & 4] Overview of molecular techniques Proteins: isolation & characterization Applications Next week: Viral model show in discussion Overview of replication/infection cycles- read Wagner,
ch 2 & 6
How do you isolate & characterize the
viruses?
Object- identify methods & reason for choicesgoal-separate from cellular proteins
sediment in ultracentrifugation; freeze; precipitate;
etc.differential centrifugation; SAS or PEG ppt;
rate zonal & density
gradient centrifugation; solvent extraction; enzyme
treatmentselectronmicrography; X-ray diffraction;
analytical ultracentrifugation; electrophoresis; ion
chromatography; serological; purity testing
Phases in infection cycle ("one-step growth curve"):1. adsorption & entry- virion NA enters cell
eukaryote V's- several hours
Nucleic acids: isolation & characterization
Genomic organization & evolution- browse
Wagner, ch 14 & Part 4, ch 3; CRC, ch 2
Updated 1/25/04 by thatcher@sonoma.edu