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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
KeywordLipomyces Starkeyi Agrobacterium-mediated Transformation Promoters Genetic Engineering Biofuels
Source PublicationAPPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
2017-11-01
DOI10.1007/s12010-017-2469-5
Volume183Issue:3Pages:867-875
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS KeywordSACCHAROMYCES-CEREVISIAE ; RHODOSPORIDIUM-TORULOIDES ; LIPID PRODUCTION ; CO-FERMENTATION ; TUMEFACIENS ; INTEGRATION ; EXPRESSION ; GENOMICS ; BIOFUELS ; CLONING
AbstractOleaginous 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.
Language英语
WOS IDWOS:000414195200013
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/149670
Collection中国科学院大连化学物理研究所
Affiliation1.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
GB/T 7714
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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