Abstract
Publisher Summary This chapter describes some of the methods for studying the synthesis and assembly of those thylakoid proteins that originate within the chloroplast. The isolation of intact chloroplasts showing high rates of protein synthesis in vitro is favored by the use of young, starch-free tissue. In the case of pea plants, excellent results are obtained by sowing seeds in moist potting fiber, allowing seedlings to develop under a 12-hr day length at day/night temperature of 22/19 o , and harvesting the leaves 9–11 days after sowing. The chapter discusses the optimum conditions for protein synthesis by isolated intact pea chloroplasts. The incorporation of L-[ 35 S]methionine into protein by isolated intact pea chloroplasts is dependent on an energy source such as light or exogenous adenosine triphosphate (ATP). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of soluble and thylakoid polypeptides, followed by autoradiography of the stained and dried gel, reveals that isolated intact chloroplasts synthesize a number of discrete translation products.
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