斑节对虾cyclin G2基因克隆及不同刺激下的表达分析
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1. 中国水产科学研究院 南海水产研究所, 农业部南海渔业资源开发利用重点实验室, 广东 广州 510300;
2. 上海海洋大学 水产与生命学院, 上海 201306;
3. 中国水产科学研究院 南海水产研究所 热带水产研究开发中心, 海南 三亚 572018;
4. 广东省海洋工程职业技术学校,

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谢波波(1990-),男,硕士研究生,从事水产养殖研究.E-mail:15521095991@163.com

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S96

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国家自然科学基金项目(31101903);海南省应用技术研发与示范推广专项(ZDXM2014057);广东省科技计划项目(2014A020208039);广东省海洋渔业科技与产业发展专项(A201501A11);农业部948计划项目(2015-Z20);海南省自然科学基金项目(20163147).


Molecular cloning and expression analysis of cyclin G2 gene from black tiger shrimps (Penaeus monodon) under different stimulation
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1. Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture; South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;
2. College of Fisheries and Life Sci

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    摘要:

    细胞周期蛋白G2(cyclin G2)是细胞周期中的重要调控因子。该研究初步探讨斑节对虾(Pmcyclin G2 cDNA全长,其开放阅读框(ORF)1161bp,编码386个氨基酸。利用荧光定量PCR技术研究了在斑节对虾的心脏、血淋巴、肝胰腺、卵巢等组织中均有表达,其中肌肉中表达量最高;卵巢发育不同阶段Pmcyclin G2基因表达升高,多巴胺(DA)则抑制卵巢中基因表达。利用原核表达技术获得了cyclin G2的体外重组蛋白,Western Blot分析证实重组蛋白为cyclin G2蛋白。此结果为进一步研究该蛋白的功能提供了条件。以上研究结果表明,基因可能与斑节对虾卵母细胞发育有密切关系,该结果可为进一步探究斑节对虾卵巢的发育机理提供理论依据。

    Abstract:

    is the largest species of shrimp, with its high economic and natural nutritional value. However, during its breeding process, broodstock breeding technology has been a very important limiting factor. With the development of molecular biology techniques, there are growing concerns about the function of cell cycle genes of crustaceans, especially shrimp. Recently, molecular mechanisms of cell cycle regulatory proteins have been researched. In the study, cell cycle-related protein family genes relating to ovarian development also become a hot topic. Cyclin G2 is a newly discovered cell cycle-related protein that plays an important role during embryonic development, cell cycle progression and development, and disease. Cyclin G2 in some species have been cloned out, but the study of the gene has not been reported in the black tiger shrimp (, mature if the eye stalk is removed. Thus, the full-length cyclin G2 cDNA sequence from ) was cloned by means of rapid amplification of cDNA ends (RACE) approaches to better understand the potential function of cyclin G2 in the regulation of shrimp reproduction. The full-length cDNA sequence was 4075 bp and contained 189 bp 5′untranslated region (UTR) and a 2725 bp 3′UTR. The open reading frame was 1161 bp and coded 386 amino acids (aa) which was highly homology to other species cyclin G2. Bioinformatics analysis showed that the amino acid coding sequence had a conserved cyclin box and the homologous protein box structure domain was 50-150 aa. The predicted molecular weight was about 43.4 kD, and the theoretical isoelectric point was 8.25. The deduced amino acid of cyclin G2 contained 29 phosphorylation sites including 17 Ser, 7 Thr, and 5 Tyr residues. SignalP 4.0 analysis revealed that cyclin G2 did not contain a typical signal peptide sequence. The temporal expression of in different tissues (ovary, heart, intestine, hepatopancreas, brain, muscles, stomach and gills) and different developmental stages of ovary was investigated by Real-time quantitative polymerase chain reaction (QRT-PCR). The lowest expression level of was observed in the hepatopancreas, and the highest in the muscle. During the ovary development stages, was significantly high expressed in stage III ovaries than the other stages. The relative expression of in the ovary was up-regulation after 5-hydroxytryptamine (5-HT) injection, while down-regulation after dopamine (DA) challenge. The study obtained recombinant expression Pmcyclin G2 in prokaryotes and offered theoretical basis for further research on Pmcyclin G2 protein function. Those results offered additional information to understand the development mechanisms of ovaries.

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谢波波,郭松,傅明骏,赵超,杨其彬,焦宗垚,邱丽华.斑节对虾cyclin G2基因克隆及不同刺激下的表达分析[J].中国水产科学,2016,23(4):823-832
XIE Bobo, GUO Song, FU Mingjun, ZHAO Chao, YANG Qibin, JIAO Zongyao, QIU Lihua. Molecular cloning and expression analysis of cyclin G2 gene from black tiger shrimps (Penaeus monodon) under different stimulation[J]. Journal of Fishery Sciences of China,2016,23(4):823-832

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  • 在线发布日期: 2016-07-21
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