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ISOLATION OF ALGAL GENES BY FUNCTIONAL COMPLEMENTATION OF YEAST 1
Author(s) -
Lippmeier J. Casey,
Brown Aaron M.,
Apt Kirk E.
Publication year - 2002
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1046/j.1529-8817.2002.01022.x
Subject(s) - biology , complementation , isolation (microbiology) , gene , computational biology , genetics , evolutionary biology , bioinformatics , phenotype
Algal cDNAs were isolated and characterized by functional complementation of yeast auxotrophs. Two cDNA libraries, one derived from the diatom Phaeodactylum tricornutum Bohlin and the other from the dinoflagellate Crypthecodinium cohnii Biecheler, were constructed using the Saccharomyces cerevisiae expression vector pFL‐61. These libraries were used for functional complementation of auxotrophic markers in two yeast strains. Yeast tryptophan auxotrophs, complemented by the P. tricornutum library, contained a plasmid that encoded a two‐domain protein associated with tryptophan synthesis, indole‐3‐glycerol phosphate synthase‐ N ‐(5′‐phosphoribosyl) anthranilate isomerase. Another cDNA originating from the C. cohnii library rescued S. cervisiae from a defect in adenine biosynthesis. This cDNA encoded a fusion of phosphoribosylamidoimidazole‐succinocarboxamide synthetase and phosphoribosylaminoimidazole carboxylase, which correspond to the yeast ade1 and ade2 genes, respectively. These results demonstrate that heterologous functional complementation can be used to identify algal genes and may provide advantages over other gene discovery methods.