Unit

UNIT 11.16 In Vitro Analysis of Chloroplast Protein Import

  1. Matthew D. Smith1,
  2. Danny J. Schnell1,
  3. Lynda Fitzpatrick2,
  4. Kenneth Keegstra2

Published Online: 1 FEB 2003

DOI: 10.1002/0471143030.cb1116s17

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Smith, M. D., Schnell, D. J., Fitzpatrick, L. and Keegstra, K. 2003. In Vitro Analysis of Chloroplast Protein Import. Current Protocols in Cell Biology. 17:11.16:11.16.1–11.16.21.

Author Information

  1. 1

    University of Massachussets, Amherst, Massachussets

  2. 2

    Michigan State University, East Lansing, Michigan

Publication History

  1. Published Online: 1 FEB 2003
  2. Published Print: JAN 2003

Abstract

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.