Localization of viral proteins in plant cells: protein tagging.
This chapter describes techniques for in vivo imaging of fluorescent fusion proteins in living cells by confocal laser scanning microscopy (CLSM). Methods are provided for (i) producing the constructs for transient expression from plasmids or virus-based vectors, (ii) introduction of constructs to plant epidermal cells; (iii) imaging of the expressed proteins by CLSM and image processing, and (iv) studying the expression in the presence of agents that affect the integrity or function of cytoskeletal elements. Notes are provided to aid comprehension and indicate problems.
Haupt
Sophie
S
University of Dundee at SCRI, Invergowrie, DD2 5DA, UK.
Ziegler
Angelika
A
Torrance
Lesley
L
D000818:Animals; D019470:Biolistics / Q000295:instrumentation / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification; D018613:Microscopy, Confocal / Q000379:methods; D010944:Plants / Q000821:virology*; D014026:Nicotiana / Q000648:ultrastructure / Q000821:virology; D014764:Viral Proteins / Q000302:isolation & purification*
D014764:Viral Proteins; D049452:Green Fluorescent Proteins