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Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides
Wang, Yanan1,3; Lin, Xinping2; Zhang, Sufang1; Sun, Wenyi1; Ma, Sijia1; Zhao, Zongbao Kent1
KeywordRhodosporidium Toruloides Promoter Transformation Genetic Engineering
Source PublicationYEAST
2016-03-01
ISSN0749-503X
DOI10.1002/yea.3145
Volume33Issue:3Pages:99-106
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Microbiology ; Mycology
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Microbiology ; Mycology
WOS KeywordSACCHAROMYCES-CEREVISIAE ; LIPID PRODUCTION ; GENE-EXPRESSION ; MULTIPLE GENES ; TRANSFORMATION ; AGROBACTERIUM ; VECTORS ; LIBRARY ; GENOME
AbstractThe oleaginous yeast Rhodosporidium toruloides is an unconventional yeast species that can accumulate a high content of lipids. Because it belongs to the basidiomycetous group of fungus, limited tools and functional elements are available for genetic engineering of R. toruloides and related red yeasts. Here we report the functional evaluation of five constitutive promoters from this yeast. We assembled a reporter gene expression cassette, consisting of a promoter, the hygromycin gene (HYG) and the nos terminator, and inserted it into the binary vector pZPK. Hygromycin-resistant transformants were obtained when R. toruloides cells were co-cultured with Agrobacterium tumefaciens AGL1 cells harbouring the engineered vector. Genomic integration of the reporter cassette was verified by successful amplification of target DNA fragments. Quantitative PCR analysis suggested that the transformant had only one copy of the reporter cassette. The strength of these promoters was demonstrated at the phenotypic level on the hygromycin-gradient plate and at the transcriptional level by real-time quantitative PCR. It was found that the strengths of these promoters varied no more than five-fold and followed a decreasing sequence of PPGI, PPGK, PFBA, PTPI, and PGPD. This study established new genetic elements for the construction of superior R. toruloides strains to produce advanced biofuels and related chemicals. Copyright (C) 2015 John Wiley & Sons, Ltd.
Language英语
WOS IDWOS:000371935200003
PublisherWILEY-BLACKWELL
Citation statistics
Cited Times:21[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/171053
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang, Sufang
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China
2.Dalian Polytech Univ, Sch Food Sci & Technol, Dalian 116034, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Wang, Yanan,Lin, Xinping,Zhang, Sufang,et al. Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides[J]. YEAST,2016,33(3):99-106.
APA Wang, Yanan,Lin, Xinping,Zhang, Sufang,Sun, Wenyi,Ma, Sijia,&Zhao, Zongbao Kent.(2016).Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides.YEAST,33(3),99-106.
MLA Wang, Yanan,et al."Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides".YEAST 33.3(2016):99-106.
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