半滑舌鳎AKT-ineracting protein基因的克隆及免疫应答表达分析

孙璐明; 于孟君; 陈亚东; 陈学杰; 刘洋; 仇雪梅; 沙珍霞

doi:10.7541/2016.62

半滑舌鳎AKT-ineracting protein基因的克隆及免疫应答表达分析

孙璐明^1,2,
于孟君^1,2,
陈亚东^2,3,
陈学杰^2,4,
刘洋¹,
仇雪梅¹,
沙珍霞^2,3, ,

1.
大连海洋大学水产与生命学院, 大连 116023
2.
中国水产科学研究院黄海水产研究所农业部海洋渔业可持续发展重点实验室, 青岛 266071
3.
青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室, 青岛 266200
4.
上海海洋大学水产与生命学院, 上海 201306

基金项目:

国家863高技术研究发展计划(2012AA10A401-4)

国家自然科学基金(31172439)资助

计量
- 文章访问数: 1344
- HTML全文浏览量: 0
- PDF下载量: 269
出版历程
- 收稿日期: 2015-09-16
- 修回日期: 2016-01-18
- 发布日期: 2016-05-24

AKT-INTERACTING PROTEIN GENE CLONING AND ITS EXPRESSION PROFILE IN RESPONSE TO PATHOGEN INFECTION IN HALF SMOOTH TONGUE SOLE (CYNOGLOSSUS SEMILAEVIS)

1.
College of Fishers and Life Sciences, Dalian Ocean University, Dalian 116023, China;
2.
Key Laboratory for Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;
3.
Laboratory for Marine Fisheries and Aquaculture, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China;
4.
College of Fishers and Life Sciences, Shanghai Ocean University, Shanghai 201306, China

Funds:

Supported by the National High Technology Research and Development Program of China (863 Program) (2012AA10A401-4)

the National Natural Science Foundation of China (31172439)

摘要

摘要: 研究克隆了半滑舌鳎Cynoglossus semilaevis膜蛋白AKT-interacting protein (AKTIP)基因, 研究了其在健康组织中的表达模式、鳗弧菌(Vibrio anguillarum)感染后免疫组织和不同病原物刺激外周血淋巴细胞中的表达特征。通过常规克隆和RACE技术, 获得的半滑舌鳎AKTIP基因全长cDNA序列为1224 bp, 其中包括5'-UTR为116 bp, 3'-UTR为117 bp和完整的ORF序列891 bp, 编码296个氨基酸, 预测蛋白质的等电点(PI)为9.12,分子量是33.74 kD; 同源比对发现半滑舌鳎AKTIP的氨基酸序列在不同物种之间具有较高的保守性; 荧光实时定量PCR (qRT-PCR)检测到半滑舌鳎各个组织中均有AKTIP基因的表达, 在卵巢中表达量最高; 鳗弧菌感染半滑舌鳎后, AKTIP基因在肝、鳃、血液、肠、头肾和脾中均上调表达; 病原模拟物PGN、LPS、poly I:C和WGP刺激半滑舌鳎外周血淋巴细胞均诱导AKTIP基因下调表达。研究表明半滑舌鳎AKTIP基因参与了机体免疫反应为给半滑舌鳎免疫防御技术的研究提供理论依据。
- 半滑舌鳎 /
- AKT-interacting protein基因 /
- 基因克隆 /
- 免疫应答 /
- 基因表达
Abstract: AKT-interacting protien (AKTIP) is a kind of membrane protein, involvimg in the regulation of PI3K/PDK1/Akt pathway. AKTIP gene is still unknown in fish. In the present study, AKTIP gene of Cynoglossus semilaevis was first cloned and full length of cDNA was 1224 bp in size, including 5'-untranslated region (UTR) of 116 bp, 3'-UTR of 117 bp and a complete open reading frame (ORF) of 891 bp, encoding 296 amino acids. Theoretical isoelectric point (PI) of predicted protein was 9.12 and molecular weight was 33.74 kD. Homologous comparison showed that the amino acids sequence of AKTIP in C. semilaevis had a high identity with those of other species. The AKTIP gene was expressed in all tested tissues in the health C. semilaevis and the highest expression was in the ovary. In order to investigate the expression patterns of the AKTIP gene in immune response, the specific expression of AKTIP was performed after Vibrio anguillarum infection and in peripheral blood lymphocytes with different pathogens stimulated. The results showed that the expression of AKTIP gene was up-regulated in the liver, gill, blood, intestinal, head kidney and spleen after V. anguillarum infection. While the AKTIP gene expression in peripheral blood lymphocytes was down-regulated after PGN, LPS, WGP and poly I: C stimulation. The research revealed that AKTIP gene involved in C. semilaevis immune response. These results provide the evidence to explore AKTIP defense mechanism and to control C. semilaevis disease in the further study.
- Cynoglossus semilaevis /
- AKT-interacting protein /
- Gene clone /
- Immune response /
- Gene expression

HTML全文

参考文献(14)

[1]	Lesche R, Peetz A, van der Hoeven F, et al. Ftl, a novel gene related to ubiquitin-conjugating enzymes, is deleted in the Fused toes mouse mutation [J]. Mammalian Genome, 1997, 8(12): 879883
[2]	Zhen S, Zhou J F, Lu L, et al. Cloning and expression pattern and over-expression analysis of Zebrafish akt3 gene [J]. Acta Hydrobiologica Sinica, 2011, 35(5): 717726 [谌爽, 周建峰, 卢玲, 等. 斑马鱼akt3基因的克隆及其表达图谱与过表达分析. 水生生物学报, 2011, 35(5): 717726]
[3]	Remy I, Michnick S W. Regulation of apoptosis by the Ft1 protein, a new modulator of protein kinase B/Akt [J]. Molecular and Cellular Biology, 2004, 24(4): 1493 1504
[4]	Woodard J, Sassano A, Hay N, et al. Statin-dependent suppression of the Akt/mammalian target of rapamycin signaling cascade and programmed cell death 4 up-regulation in renal cell carcinoma [J]. Clinical Cancer Research, 2008, 14(14): 46404649
[5]	Jiang H, Zhang X J. PKB/Akt: A protein kinase that has a variety of functions [J]. Life Science, 2004, 16(3): 148153 [姜华, 张学军. PKB/Akt: 一个具有多种功能的蛋白激酶. 生命科学, 2004, 16(3): 148153]
[6]	Romashkova J A, Makarov S S. NF-B is a target of AKT in anti-apoptotic PDGF signaling [J]. Nature, 1999, 401(6748): 8690
[7]	Sha Z X, Wang Q L, Liu Y, et al. Identification and expression analysis of goose-type lysozyme in half-smooth tongue sole (Cynoglossus semilaevis) [J]. Fish Shellfish Immunology, 2012, 32(5): 914921
[8]	Sha Z, Gong G, Wang S, et al. Identification and characterization of Cynoglossus semilaevis microRNA response to Vibrio anguillarum infection through high-throughput sequencing [J]. Developmental Comparative Immunology, 2014, 44(1): 5969
[9]	Fatrai S, Elghazi L, Balcazar N, et al. Akt induces -cell proliferation by regulating cyclin D1, cyclin D2, and p21 levels and cyclin-dependent kinase-4 activity [J]. Diabetes, 2006, 55(2): 318325
[10]	Datta S R, Dudek H, Tao X, et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery [J]. Cell, 1997, 91(2): 231241
[11]	Nicholson K M, Anderson N G. The protein kinase B/Akt signalling pathway in human malignancy [J]. Cellular signalling, 2002, 14(5): 381395
[12]	Yamada T, Takeuchi S, Fujita N, et al. Akt kinase-interacting protein1, a novel therapeutic target for lung cancer with EGFR-activating and gatekeeper mutations [J]. Oncogene, 2013, 32(37): 44274435
[13]	Chandrasekar B, Mummidi S, Perla R, et al. Fractalkine (CX3CL1) stimulated by nuclear factor kappaB (NF-kappaB)-dependent inflammatory signals induces aortic smooth muscle cell proliferation through an autocrine pathway [J]. Biochemical Journal, 2003, 373: 547558
[14]	Nakamura A, Naito M, Tsuruo T, et al. Freud-1/Aki1, a novel PDK1-interacting protein, functions as a scaffold to activate the PDK1/Akt pathway in epidermal growth factor signaling [J]. Molecular and Cellular Biology, 2008, 28(19): 59966009

施引文献(7)

期刊类型引用(3)

1.	李兆龙，王腾腾，韩慧宗，陈钰臻，王斐，张明亮，孙硕，解维俊，姜海滨. 许氏平鲉蛋白激酶B(SsAkt)基因的克隆及其mRNA在细菌胁迫后的表达规律. 渔业科学进展. 2024(01): 47-59 . 百度学术
2.	马慧鑫，王磊，汪林庆，周茜，陈松林. 牙鲆精氨酸酶Ⅱ基因的克隆以及免疫应答表达分析. 渔业科学进展. 2018(03): 44-52 . 百度学术
3.	汪林庆，王航，陈亚东，杨光，白莉，孙璐明，沙珍霞. 半滑舌鳎外周血T淋巴细胞的分离、鉴定及TCRβ基因免疫应答分析. 水生生物学报. 2018(03): 480-487 . 本站查看

其他类型引用(4)

资源附件(0)

计量

文章访问数: 1344
HTML全文浏览量: 0
PDF下载量: 269
被引次数: 7

半滑舌鳎AKT-ineracting protein基因的克隆及免疫应答表达分析

计量

出版历程

AKT-INTERACTING PROTEIN GENE CLONING AND ITS EXPRESSION PROFILE IN RESPONSE TO PATHOGEN INFECTION IN HALF SMOOTH TONGUE SOLE (CYNOGLOSSUS SEMILAEVIS)

期刊类型引用(3)

其他类型引用(4)

计量

出版历程

目录