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Isolation and characterization of the TRP1 gene from the yeast Yarrowia lipolytica and multiple gene disruption using a TRP blaster
Author(s) -
Cheon Seon Ah,
Han Eun Jung,
Kang Hyun Ah,
Ogrydziak David M.,
Kim JeongYoon
Publication year - 2003
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.987
Subject(s) - yarrowia , biology , complementation , gene , saccharomyces cerevisiae , genetics , yeast , gene cluster , kluyveromyces lactis , genbank , nucleic acid sequence , sequence analysis , accession number (library science) , mutant
The TRP1 gene encoding N‐(5′‐phosphoribosyl)‐anthranilate isomerase was isolated from the yeast Yarrowia lipolytica , in which only a few genetic marker genes are available. The Y. lipolytica TRP1 gene ( YlTRP1 ) cloned by complementation of Y. lipolytica trp1 mutation was found to be a functional homologue of Saccharomyces cerevisiae TRP1 . Since YlTRP1 could be used for counterselection in medium containing 5‐fluoroanthranilic acid (5‐FAA), we constructed TRP blasters that contained YlTRP1 flanked by a direct repeat of a sequence and allowed the recycling of the YlTRP1 marker. Using the TRP blasters the sequential disruption of target genes could be carried out within the same strain of Y. lipolytica . The nucleotide sequence of the YlTRP1 gene has been deposited at GenBank under Accession No. AF420590. Copyright © 2003 John Wiley & Sons, Ltd.