In vitro analysis of chloroplast protein import.
This unit describes protocols for isolating chloroplasts from pea (Pisum sativum) and Arabidopsis thaliana for the study of nuclear-encoded plastid precursor proteins. Chloroplasts from both preparations are competent for the in vitro import of recombinant preproteins synthesized using in vitro translation systems derived from reticulocyte or wheat germ lysates. These assays can be used to test whether a particular protein is targeted to chloroplasts, for analyzing the suborganellar location of newly imported preproteins, or to study the mechanism of import itself.
Smith
Matthew D
MD
University of Massachussets, Amherst, Massachussets, USA.
Schnell
Danny J
DJ
Fitzpatrick
Lynda
L
Keegstra
Kenneth
K
D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods; D002736:Chloroplasts / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D018532:Pisum sativum / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis* / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects
D010940:Plant Proteins