Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides

doi:10.1002/yea.3145

	Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides
	Wang, Yanan 1,3; Lin, Xinping 2; Zhang, Sufang 1; Sun, Wenyi 1; Ma, Sijia 1; Zhao, Zongbao Kent 1
刊名	YEAST
	2016-03-01
卷号	33 期号:3 页码:99-106
关键词	Rhodosporidium Toruloides Promoter Transformation Genetic Engineering
ISSN号	0749-503X
DOI	10.1002/yea.3145
文献子类	Article
英文摘要	The 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.
WOS关键词	SACCHAROMYCES-CEREVISIAE ; LIPID PRODUCTION ; GENE-EXPRESSION ; MULTIPLE GENES ; TRANSFORMATION ; AGROBACTERIUM ; VECTORS ; LIBRARY ; GENOME
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Microbiology ; Mycology
语种	英语
出版者	WILEY-BLACKWELL
WOS记录号	WOS:000371935200003
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/171052]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Zhang, Sufang
作者单位	1.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
推荐引用方式 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.