Cloning and characterization of a novel apolipoprotein gene, apolipoprotein AV, in tree shrews
Cloning and characterization of a novel apolipoprotein gene, apolipoprotein AV, in tree shrews
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树鼩新型载脂蛋白基因载脂蛋白 AV 的克隆和表征
DOI：
10.1007/s11033-013-2641-0
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发表时间：
2013-05
影响因子：
2.8
通讯作者：
Chen, Baosheng
中科院分区：
文献类型：
--
作者：
Man, Yong;Wang, Shu;Li, Jian;Chen, Baosheng
载脂蛋白AV（apoAV）调节血浆甘油三酯水平，这是一个独立的危险因素，心血管疾病。ApoAV还参与动脉粥样硬化病变形成。为了系统地评估树鼠（一种动脉粥样硬化不敏感模型）的载脂蛋白相关基因谱，我们进行了apoAV克隆和鉴定。采用SMART-RACE技术对apoAV的全长cDNA进行鉴定。ApoAV cDNA序列显示两个转录本，1，948和1，397碱基对，由于交替聚腺苷酸化。这两个转录本共享相同的开放阅读框架（ORF），其编码369个氨基酸的蛋白，与人apoAV具有高度同源性（75%），包括23个氨基酸的N-末端信号肽。ApoAV仅在肝脏中表达。成熟的apoAV在E. coliBL 21（DE 3），镍螯合树脂纯化。该重组蛋白对脂蛋白脂酶活性有明显的刺激作用。将apoAV的全长ORF克隆到带有红色荧光蛋白标签的pDsRed-monomer-N1载体中，并初步定位于hepG 2细胞的胞浆中。apoAV基因在树鼩中的成功克隆、表达和定位为进一步研究其结构和功能奠定了基础。
Apolipoprotein AV (apoAV) modulates plasma triglyceride levels, which is an independent risk factor for cardiovascular disease. ApoAV is also involved in atherosclerosis lesion formation. In order to systematically evaluate the apolipoprotein-related gene profile in tree shrew, a model for its insusceptibility to atherosclerosis, we performed apoAV cloning and characterization. The full-length cDNA of apoAV was identified using SMART-RACE. ApoAV cDNA sequence revealed two transcripts, 1,948 and 1,397 base pairs, due to alternative polyadenylation. These two transcripts share the same open reading frame (ORF), which encodes a 369-amino acid protein with high identity to human apoAV (75 %), including a 23-amino acid N-terminal signal peptide. ApoAV is expressed exclusively in the liver. Mature apoAV was expressed inE. coliBL21(DE3) and purified by Ni-chelated resin. Lipoprotein lipase activity was significantly stimulated by this recombinant protein. The full-length ORF of apoAV was cloned into pDsRed-monomer-N1 vector with a red fluorescent protein tag and was primarily localized in cytoplasm of hepG2 cells. The successful cloning, expression and localization of apoAV in tree shrew has laid down the foundation for further investigation on its structure and functions.
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影响因子：
3.5
作者：
Pennacchio, LA;Olivier, M;Cohen, JC
通讯作者：
Cohen, JC
影响因子：
5.3
作者：
Endo, K;Yanagi, H;Tomura, S
通讯作者：
Tomura, S
影响因子：
2.8
作者：
W. Zeng;B. Zhou;Hui-rong Liu;Guotao Sun;Bao-sheng Chen
通讯作者：
W. Zeng;B. Zhou;Hui-rong Liu;Guotao Sun;Bao-sheng Chen
影响因子：
6.1
作者：
W. Zeng;Jian Zhang;Bao-sheng Chen;Gang Wu;H. Xue
通讯作者：
W. Zeng;Jian Zhang;Bao-sheng Chen;Gang Wu;H. Xue
影响因子：
4.7
作者：
Evans, D;Buchwald, A;Beil, FU
通讯作者：
Beil, FU