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学科主题微生物学
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)
会议文集Book of Abstracts
会议名称Asian Congress on Biotechnology
会议日期2011-5-11
2011
会议地点上海
页码58-0
出版者待补充
出版地待补充
合作性质分会口头报告
部门归属1816
主办者亚洲生物技术联合会
英文摘要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.; 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.
语种英语
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/116039
专题中国科学院大连化学物理研究所
通讯作者Zhao ZB(赵宗保)
推荐引用方式
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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