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Rapid purification of intact chloroplasts and heat-bleached ribosome-deficient plastids from rye leaves on discontinuous PercollTM gradients

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Summary

The distribution of the major cellular organelles from normal green and heat-bleached 70S ribosome-deficient rye leaves was analyzed by the assay of marker enzymes after isopycnic centrifugation on linear Percoll gradients. A rapid procedure with stepped Percoll gradients was developed for the purification of intact plastids, virtually free of contaminations by mitochondria and peroxisomes. The intact plastids are separated from broken plastids by a density step and from mitochondria and microbodies by their higher sedimentation rate. In the presence of appropriate additives in the grinding and gradient media Percoll-gradient-purified intact rye chloroplasts were capable of high rates of bicarbonate-dependent O2-evolution.

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Abbreviations

BSA:

bovine serum albumin

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

PEG:

polyethylene glycol

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Authors and Affiliations

  1. Botanisches Institut, J. W. Goethe-UniversitÄt, Postfach 111932, D-6000, Frankfurt/M. 11, Federal Republic of Germany

    R. Höinghaus & J. Feierabend

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  1. R. Höinghaus
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  2. J. Feierabend
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Höinghaus, R., Feierabend, J. Rapid purification of intact chloroplasts and heat-bleached ribosome-deficient plastids from rye leaves on discontinuous PercollTM gradients. Protoplasma 118, 114–120 (1983). https://doi.org/10.1007/BF01293067

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  • DOI: https://doi.org/10.1007/BF01293067

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