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题名: Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi
作者: Lin, Xinping1;  Liu, Sasa1;  Bao, Ruiqi1;  Gao, Ning1;  Zhang, Sufang2;  Zhu, Rongqian1;  Zhao, Zongbao Kent2
关键词: Lipomyces starkeyi ;  Agrobacterium-mediated transformation ;  Promoters ;  Genetic engineering ;  Biofuels
刊名: APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
发表日期: 2017-11-01
DOI: 10.1007/s12010-017-2469-5
卷: 183, 期:3, 页:867-875
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology
研究领域[WOS]: Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology
英文摘要: Oleaginous yeast Lipomyces starkeyi, a promising strain of great biotechnical importance, is able to accumulate over 60% of its cell biomass as triacylglycerols (TAGs). It is promising to directly produce the derivatives of TAGs, such as long-chain fatty acid methyl esters and alkanes, in L. starkeyi. However, techniques for genetic modification of this oleaginous yeast are lacking, thus, further research is needed to develop genetic tools and functional elements. Here, we used two exogenous promoters (pGPD and pPGK) from oleaginous yeast Rhodosporidium toruloides to establish a simpler Agrobacterium-mediated transformation (AMT) method for L. starkeyi. Hygromycin-resistant transformants were obtained on antibiotic-contained plate. Mitotic stability test, genotype verification by PCR, and protein expression confirmation all demonstrated the success of this method. Furthermore, the strength of these two promoters was evaluated at the phenotypic level on a hygromycin-gradient plate and at the transcriptional level by real-time quantitative PCR. The PGK promoter strength was 2.2-fold as that of GPD promoter to initiate the expression of the hygromycin-resistance gene. This study provided an easy and efficient genetic manipulation method and elements of the oleaginous yeast L. starkeyi for constructing superior strains to produce advanced biofuels.
关键词[WOS]: SACCHAROMYCES-CEREVISIAE ;  RHODOSPORIDIUM-TORULOIDES ;  LIPID PRODUCTION ;  CO-FERMENTATION ;  TUMEFACIENS ;  INTEGRATION ;  EXPRESSION ;  GENOMICS ;  BIOFUELS ;  CLONING
语种: 英语
WOS记录号: WOS:000414195200013
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/149670
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Dalian Polytech Univ, Sch Food Sci & Technol, Natl Engn Res Ctr Seafood, Dalian 116034, Peoples R China
2.Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China

Recommended Citation:
Lin, Xinping,Liu, Sasa,Bao, Ruiqi,et al. Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi[J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,2017,183(3):867-875.
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