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BIBTEX RIS

Simplified Procedure for the Isolation of Intact Chloroplasts from Chlamydomonas reinhardtii

1991; Oxford University Press; Volume: 97; Issue: 4 Linguagem: Inglês

10.1104/pp.97.4.1576

ISSN

1532-2548

Autores

Catherine Mason, Sharon W. Matthews, Terry Bricker, James V. Moroney,

Tópico(s)

Photoreceptor and optogenetics research

Resumo

A simple procedure that yields highly purified intact chloroplasts from Chlamydomonas reinhardtii is described. This procedure involves breakage of cell wall-deficient cells by passing them through a narrow bore syringe needle. The intact chloroplasts are then purified from the crude homogenate by differential centrifugation and Percoll gradient centrifugation. This procedure generates relatively high yields of chloroplasts capable of CO(2) fixation. These chloroplasts were characterized by electron microscopy, marker enzyme analysis, and ferricyanide exclusion. Transmission electron microscopy indicates that these chloroplasts retain their pyrenoids and eyespots. Scanning electron microscopy confirms that the characteristic cup shape of C. reinhardtii chloroplasts persists in vitro. This rapid, inexpensive procedure produces chloroplasts that should be useful for researchers studying the biochemistry and cell biology of C. reinhardtii chloroplasts.

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