Functional genomics of eukaryotic photosynthesis using insertional mutagenesis of Chlamydomonas reinhardtii.

Abstract:

:The unicellular green alga Chlamydomonas reinhardtii is a widely used model organism for studies of oxygenic photosynthesis in eukaryotes. Here we describe the development of a resource for functional genomics of photosynthesis using insertional mutagenesis of the Chlamydomonas nuclear genome. Chlamydomonas cells were transformed with either of two plasmids conferring zeocin resistance, and insertional mutants were selected in the dark on acetate-containing medium to recover light-sensitive and nonphotosynthetic mutants. The population of insertional mutants was subjected to a battery of primary and secondary phenotypic screens to identify photosynthesis-related mutants that were pigment deficient, light sensitive, nonphotosynthetic, or hypersensitive to reactive oxygen species. Approximately 9% of the insertional mutants exhibited 1 or more of these phenotypes. Molecular analysis showed that each mutant line contains an average of 1.4 insertions, and genetic analysis indicated that approximately 50% of the mutations are tagged by the transforming DNA. Flanking DNA was isolated from the mutants, and sequence data for the insertion sites in 50 mutants are presented and discussed.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Dent RM,Haglund CM,Chin BL,Kobayashi MC,Niyogi KK

doi

10.1104/pp.104.055244

subject

Has Abstract

pub_date

2005-02-01 00:00:00

pages

545-56

issue

2

eissn

0032-0889

issn

1532-2548

pii

pp.104.055244

journal_volume

137

pub_type

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