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Characterization of Carotenogenesis Genes in the Cyanobacterium Anabaena sp. PCC 7120

 Last Update: 2021-02-24
 Source: Internet
 Author: User

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Cyanobacteria produce many kinds of carotenoids for light harvesting and light protection in photosynthesis. To elucidate the biosynthetic pathways of carotenoids in
Anabaena
sp. PCC 7120 (also known as
Nostoc
sp. PCC 7120), we have produced gene-disruption mutants lacking selected proposed carotenoid biosynthetic enzymes. Here we describe the construction of mutants by triparental mating. A cargo plasmid, bearing a target gene interrupted by an antibiotic-resistant cassette, is transformed to
E. coli
donor containing a helper plasmid, and is introduced into
Anabaena
cells by conjugation. Double-reciprocal recombination replaces the target genes in
Anabaena
genome with mutated ones on the plasmid. Carotenoids in the selected double recombinants are identified using high-performance liquid chromatography.

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