Bulk Segregant Analysis by High-Throughput Sequencing Reveals a Novel Xylose Utilization Gene from Saccharomyces cerevisiae 英文参考文献.docVIP
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Bulk Segregant Analysis by High-Throughput Sequencing Reveals a Novel Xylose Utilization Gene from Saccharomyces cerevisiae 英文参考文献
BulkSegregantAnalysisbyHigh-Throughput
SequencingRevealsaNovelXyloseUtilizationGenefrom
Saccharomycescerevisiae
JaredW.Wenger,KatjaSchwartz,GavinSherlock*
DepartmentofGenetics,StanfordUniversity,Stanford,California,UnitedStatesofAmerica
Abstract
Fermentationofxyloseisafundamentalrequirementfortheefficientproductionofethanolfromlignocellulosicbiomass
sources.Althoughtheyaggressivelyfermenthexoses,ithaslongbeenthoughtthatnativeSaccharomycescerevisiaestrains
cannotgrowfermentativelyornon-fermentativelyonxylose.Populationsurveyshaveuncoveredafewnaturallyoccurring
strainsthatareweaklyxylose-positive,andsomeS.cerevisiaehavebeengeneticallyengineeredtofermentxylose,butno
strain, either natural or engineered, has yet been reported to ferment xylose as efficiently as glucose. Here, we used a
medium-throughputscreentoidentifySaccharomycesstrainsthatcanincreaseinopticaldensitywhenxyloseispresented
as the sole carbon source. We identified 38 strains that have this xylose utilization phenotype, including strains of S.
cerevisiae,othersensustrictomembers,andhybridsbetweenthem.AlltheS.cerevisiaexylose-utilizingstrainsweidentified
arewineyeasts,andforthosethatcouldproducemeioticprogeny,thexylosephenotypesegregatesasasinglegenetrait.
WemappedthisgenebyBulkSegregantAnalysis(BSA)usingtilingmicroarraysandhigh-throughputsequencing.Thegene
isaputativexylitoldehydrogenase,whichwenameXDH1,andislocatedinthesubtelomericregionoftherightendof
chromosomeXVinaregionnotpresentintheS288creferencegenome.Wefurthercharacterizedthexylosephenotypeby
performinggeneexpressionmicroarraysandbygeneticallydissectingtheendogenousSaccharomycesxylosepathway.We
havedemonstratedthatnaturalS.cerevisiaeyeastsarecapableofutilizingxyloseasthesolecarbonsource,characterized
thegeneticbasisforthistraitaswellastheendogenousxyloseutilizationpathway,anddemonstratedthefeasibilityofBSA
usinghigh-throughputsequencing.
Citation: Wenger JW, Schwartz K, Sherlock G (2010) Bulk Segregant Analysis by High-Throughput Sequencing Re
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