斑节对虾细胞周期蛋白E基因的克隆与表达分析

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斑节对虾细胞周期蛋白E基因的克隆与表达分析
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作者单位:1. 中国水产科学研究院 南海水产研究所, 广东 广州510300;2. 上海海洋大学 水产与生命学院, 上海2013063. 农业部南海渔业资源开发利用重点实验室, 广东 广州510300
作者简介:赵超(1987-), 硕士, 专业方向为水产养殖. E-mail: zhaochao@126.com
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中图分类号:S917
基金项目:国家863计划项目(2012AA10A409); 国家自然科学基金项目(40976101).

Molecular cloning and expression analysis of the cyclin E gene from black tiger shrimp Penaeus monodon

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1. South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;2. College of Aqua-life Science and Technology, Shanghai Ocean University, Shanghai 201306, China;3. Key Laboratory of South China Sea Fishery Re

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发育过程中的作用RACE1 706 bpPmcyclin E1 263 bp个氨基酸。生物信息学分析表明CYCLIN该基因编码蛋白与红火蚁等多种节肢动物细胞周期蛋白其中卵巢中表达量较高(MIH)后细胞

Abstract:

Cyclins are the positive regulatory subunits of cyclin-dependent kinases (CDK). They play a critical role in the coordination of the eucaryotic cell cycle by binding to the catalytic subunits of CDKs. They share distant sequence homology over a 100 amino acid region called the cyclin box and have been classified into two phylogenetically divergent subfamilies. The first contains A, B, D, E, F, and G cyclins and the second contains the C and H cyclins. Shrimp culture is limited by a lack of methods to breed adults in captivity. This is because female shrimp are unable to reach ovarian maturity without intervention using destructive, expensive, or time consuming methods such as eyestalk-ablation. Thus, there is a need to better understand the of ovarian development. Cyclin E is a cell cycle regulatory factor that plays a very important role in cell proliferation. To better understand the potential function of cell cyclin E in regulating shrimp reproduction, the full-length cDNA sequence of cell cyclin E from black tiger shrimps (denoted as Pmcyclin E) was obtained by high throughput transcriptome sequencing and RACE. The cDNA of Pmcyclin E was 1 706 bp, including a 132 bp 5′-terminal un-translated region (cUTR), a 311 bp 3′ UTR with a poly (A) tail, and an open reading frame (ORF) of 1 263 bp encoding a polypeptide of 420 amino acids with a predicted molecular weight of 47.9 kD and predicted pI of 5.85. The results of Blast and phylogenetic analyses suggest that Pmcyclin E is a new member of the shrimp cell cyclin E family. We measured mRNA expression of Pmcyclin E in nine tissues by real-time PCR. Expression was highest in the ovary and eyestalk, moderate in the intestine, brain, and muscle, and very low in the stomach and . Pmcyclin E expression in the ovary was significantly reduced by injection of MIH, and was induced after eyestalk ablation, suggesting that the gene plays an important role in Our results provide insight into the function of Pmcyclin E in the regulation of ovarian development in the black tiger shrimp.

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赵超,傅明骏,江世贵,周发林,杨其彬,朱彩艳,邱丽华.斑节对虾细胞周期蛋白E基因的克隆与表达分析[J].中国水产科学,2014,21(3):464-473
ZHAO Chao, FU Mingjun, JIANG Shigui, ZHOU Falin, YANG Qibin, ZHU Caiyan, QIU Lihua. Molecular cloning and expression analysis of the cyclin E gene from black tiger shrimp Penaeus monodon[J]. Journal of Fishery Sciences of China,2014,21(3):464-473

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