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学科主题: 物理化学
题名: Cloning and characterization of the isocitrate dehydrogenase gene from oleaginous yeast Lipomyces starkeyi
作者: Tang W(唐伟) ;  Zhang SF(张素芳) ;  Tan HD(谭海东) ;  Zhao ZB(赵宗保)
会议名称: 13th International Biotechnology Symposium and Exhibition
会议日期: 2008-10-12
出版日期: 2008-10-12
会议地点: 中国
通讯作者: 赵宗保
ISSN: 0168-1656
部门归属: 十八室
主办者: 中国科学院,中国工程院,中国教育部,中国科技部
摘要: Oleaginous yeast Lipomyces starkeyi can accumulate intracellular lipids over 65% of its cell dry weight [1]. Mitochondrial NAD+-specific isocitrate dehydrogenase (IDH) catalyzes the oxidative decarboxylation of isocitrare to α-ketoglutarate in eukaryotic cells, an essential and rate-limiting step in citric acid cycle. Yeast S. cerevisiae IDH is composed of an octamer of four IDH1 and four IDH2 subunits [2]. Early biochemical study showed that IDH activity correlates with the lipid storage capacity [3]. In this work we cloned the full length cDNA sequence of the mitochondrial IDH1 and IDH2 gene using methods of degenerate polymerase chain reaction (PCR) and rapid amplification of cDNA ends from L. starkeyi AS 2.1560, a useful oil-producing yeast. We further determined the expression level of both lsidh1 and lsidh2 by real time PCR during the lipid storage process. Our results showed the encoded products, LsIDH1 and LsIDH2, are similar in size with a molecular weight of 40,277 and 40,708, respectively, and have high similarity with IDH sequences from other yeast species. More importantly, putative mitochondrial transit peptides have been identified at the N-terminus of each sequence, suggesting that both proteins are likely located within the mitochondria. Further gene expression analysis by real time PCR showed that both idh1 and idh2 expression decreased concurrently with the evolution of cellular lipids. Our data may be valuable for further engineering oleaginous yeasts.
语种: 中文
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内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/113180
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Tang W,Zhang SF,Tan HD,et al. Cloning and characterization of the isocitrate dehydrogenase gene from oleaginous yeast Lipomyces starkeyi[C]. 见:13th International Biotechnology Symposium and Exhibition. 中国. 2008-10-12.
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