DICP OpenIR
Subject Area微生物学
Deletion of the DGAAT gene with a bleomycin resistance cassette in the oleaginous yeast Rhodosporidium toruloides
Lin XP(林心萍); Zhang SF(张素芳); Yang F(杨帆); Zhu ZW(朱志伟); Zhao ZB(赵宗保)
Source PublicationBook of Abstracts
Conference NameAsian Congress on Biotechnology
Conference Date2011-5-11
2011
Conference Place上海
Pages58-0
Publisher待补充
Publication Place待补充
Cooperation Status分会口头报告
Department1816
Funding Organization亚洲生物技术联合会
AbstractThe red yeast Rhodosporidium toruloides is oleaginous with a great propensity to accumulate intracellular lipids over 70% of its dry cell weight when carbohydrates were fed as carbon sources. Therefore, this yeast has been intensively studied recently in order to produce microbial lipid for a sustainable biodiesel industry. On the other hand, the biochemical machinery of lipid metabolism may be explored for production of value-added metabolites. For example, diacylglycerols, the precursor to triacylglycerols, are attractive because of their special biological activities. In this study, we isolated the genomic DNA sequence of the acyl-coenzyme A:diacylglycerol acyltransferase gene (DGAAT). We generated a knockout construction with bleomycin resistance cassette replacing the 1858 bp open reading frame of the DGAAT gene. The haploid strain R. toruloides NP11 was electroporated with the cassette, and transformant was observed on YEPD medium containing 50 μg/ml Zeocine. It should be noted that the wild-type strain were killed by 5 μg/ml Zeocine. Further PCR analysis and genome walking showed that the bleomycin cassette was integrated into the chromosome of the yeast. These results demonstrated that cassette was functional in R. toruloides under the control of the DGAAT promoter and the terminator. We will discuss more details and progress of this project during the conference.; The red yeast Rhodosporidium toruloides is oleaginous with a great propensity to accumulate intracellular lipids over 70% of its dry cell weight when carbohydrates were fed as carbon sources. Therefore, this yeast has been intensively studied recently in order to produce microbial lipid for a sustainable biodiesel industry. On the other hand, the biochemical machinery of lipid metabolism may be explored for production of value-added metabolites. For example, diacylglycerols, the precursor to triacylglycerols, are attractive because of their special biological activities. In this study, we isolated the genomic DNA sequence of the acyl-coenzyme A:diacylglycerol acyltransferase gene (DGAAT). We generated a knockout construction with bleomycin resistance cassette replacing the 1858 bp open reading frame of the DGAAT gene. The haploid strain R. toruloides NP11 was electroporated with the cassette, and transformant was observed on YEPD medium containing 50 μg/ml Zeocine. It should be noted that the wild-type strain were killed by 5 μg/ml Zeocine. Further PCR analysis and genome walking showed that the bleomycin cassette was integrated into the chromosome of the yeast. These results demonstrated that cassette was functional in R. toruloides under the control of the DGAAT promoter and the terminator. We will discuss more details and progress of this project during the conference.
Language英语
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/116039
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhao ZB(赵宗保)
Recommended Citation
GB/T 7714
Lin XP,Zhang SF,Yang F,et al. Deletion of the DGAAT gene with a bleomycin resistance cassette in the oleaginous yeast Rhodosporidium toruloides[C]. 待补充:待补充,2011:58-0.
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