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Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi
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
2017-11-01
DOI10.1007/s12010-017-2469-5
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
关键词[WOS]SACCHAROMYCES-CEREVISIAE ; RHODOSPORIDIUM-TORULOIDES ; LIPID PRODUCTION ; CO-FERMENTATION ; TUMEFACIENS ; INTEGRATION ; EXPRESSION ; GENOMICS ; BIOFUELS ; CLONING
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.
语种英语
WOS记录号WOS:000414195200013
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被引频次:2[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/149733
专题中国科学院大连化学物理研究所
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
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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.
APA Lin, Xinping.,Liu, Sasa.,Bao, Ruiqi.,Gao, Ning.,Zhang, Sufang.,...&Zhao, Zongbao Kent.(2017).Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi.APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,183(3),867-875.
MLA Lin, Xinping,et al."Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi".APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 183.3(2017):867-875.
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