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The protein complexes in the thylakoid membrane are composed of subunits derived from both the nuclear and chloroplast genomes. While less is known about the mechanisms of delivery of the plastid-encoded subunits, the targeting mechanisms of the nuclear-encoded subunits have been more experimentally tractable. We have described in this chapter the methods used in our laboratory for investigations of the import of nuclear-encoded proteins across the chloroplast envelope membranes, and for their further delivery into or across the thylakoid membrane by one of the four distinct pathways.