水牛浮舰蛋白2基因克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2016.04.009

›› 2016, Vol. 43 ›› Issue (4): 913-920.doi: 10.16431/j.cnki.1671-7236.2016.04.009

水牛浮舰蛋白2基因克隆及序列分析

段安琴, 庞春英, 朱鹏, 邓廷贤, 陆杏蓉, 梁贤威

中国农业科学院广西水牛研究所, 广西水牛遗传繁育重点实验室, 南宁 530001

收稿日期:2015-10-10 出版日期:2016-04-20 发布日期:2016-04-27
通讯作者: 梁贤威 E-mail:liangbri@126.com
作者简介:段安琴(1988-),女,广西柳州人,研究实习员,硕士生,研究方向:动物生殖生理,E-mail:duanaq321@163.com
基金资助:
农业部转基因重点项目(2014ZX0801012B);桂科合(14123001-5);水牛基(1504005)

Cloning and Sequence Analysis of Buffalo FLOT2 Gene

DUAN An-qin, PANG Chun-ying, ZHU Peng, DENG Ting-xian, LU Xing-rong, LIANG Xian-wei

Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Science, Nanning 530001, China

Received:2015-10-10 Online:2016-04-20 Published:2016-04-27

摘要/Abstract

摘要： 试验旨在克隆水牛浮舰蛋白2(Flotillin 2,FLOT2)基因并对其进行生物信息学分析,为揭示该基因在水牛发育繁殖中的作用奠定基础。本研究通过PCR扩增获得了FLOT2基因全长并进行克隆测序,结合生物信息学分析方法,预测及分析其蛋白质结构。结果表明,水牛FLOT2基因编码区长1 287 bp,编码428个氨基酸,3'UTR区长277 bp。多重序列比较分析显示,水牛FLOT2基因核苷酸序列与牛、山羊、绵羊、小鼠、马、猫和人相应序列的相似性分别为98%、98%、97%、90%、93%、92%和92%,系统进化树分析结果表明,FLOT2基因在不同物种以及进化的过程中具有高度保守性。对FLOT2蛋白质的分析表明,该蛋白呈弱酸性,无信号肽,细胞亚定位于细胞质中,存在SPFH_flotillin和SPFH_hflk等结构域。microRNA预测结果表明,bta-miR-2438、bta-miR-2379和bta-miR-1777a可能是其3'UTR潜在靶标。研究结果为今后阐明FLOT2基因在水牛繁殖性能中的功能,尤其是在配子及胚胎发生过程中的作用及分子机制奠定了基础。

关键词: 水牛; 浮舰蛋白2基因; 克隆; 分析

Abstract: In order to clarify the effect of Flotillin 2 (FLOT2) gene on the reproductive performance of buffalo, buffalo FLOT2 gene was cloned and analyzed by bioinformatics.FLOT2 was cloned by PCR, cloned and sequenced, then its protein structure was predicted and analyzed by bioinformatics tools.The results showed that the coding region of buffalo FLOT2 gene was 1 287 bp, 3'UTR was 277 bp, encoded 428 amino acids.The buffalo FLOT2 gene shared 98%, 98%, 97%, 90%, 93%, 92% and 92% of similar nucleotide sequence with that of Bos taurus, Ovis aries, Capra hircus, Mus musculus, Equus caballus, Felis catus and Homo sapiens, respectively.Phylogenetic tree analysis showed that FLOT2 gene was highly conserved in different species and evolution.FLOT2 protein was weakly acidic, without signal peptide, located in the cytoplasmic, and with the presence of SPFH_flotillin and SPFH_hflk domain.microRNA prediction showed bta-miR-2438, bta-miR-2379 and bta-miR-1777a maybe target the 3'UTR of buffalo FLOT2.In a word, this study provided an important reference for surveying the regulation mechanism of FLOT2 gene in buffalo reproductive performance, especially, during buffalo gametogenesis and embryogenesis.

Key words: buffalo; FLOT2 gene; cloning; analysis

中图分类号:

Q785

段安琴, 庞春英, 朱鹏, 邓廷贤, 陆杏蓉, 梁贤威. 水牛浮舰蛋白2基因克隆及序列分析[J]. , 2016, 43(4): 913-920.

DUAN An-qin, PANG Chun-ying, ZHU Peng, DENG Ting-xian, LU Xing-rong, LIANG Xian-wei. Cloning and Sequence Analysis of Buffalo FLOT2 Gene[J]. , 2016, 43(4): 913-920.

参考文献

[1] Allen J A,Halverson-Tamboli R A,Rasenick M M.Lipid raft microdomains and neurotransmitter signalling[J].Nat Rev Neurosci,2007,8(2):128-140.
[2] Schulte T,Paschke K A,Laessing U,et al.Reggie-1 and Reggie-2,two cell surface proteins expressed by retinal ganglion cells during axon regeneration[J]. Development,1997,124(2):577-587.
[3] Bickel P E,Scherer P E,Schnitzer J E,et al.Flotillin and epidermal surface antigen define a new family of caveolae-associated integral membrane proteins[J].J Biol Chem,1997,272(21):13793-13802.
[4] Cao K,Xie D,Cao P,et al.SiRNA-mediated flotillin-2(FLOT2) downregulation inhibits cell proliferation,migration,and invasion in gastric carcinoma cells[J]. Oncol Res,2014,21(5):271-279.
[5] Zhao F,Zhang J,Liu Y S,et al.Research advances on flotillins[J]. Virol J,2011,8:479.
[6] Gomez V,Sese M,Santamaria A,et al.Regulation of aurora B kinase by the lipid raft protein flotillin-1[J].J Biol Chem,2010,285(27):20683-20690.
[7] Langhorst M F,Reuter A,Luxenhofer G,et al.Preformed reggie/flotillin caps:Stable priming platforms for macrodomain assembly in T cells[J]. Faseb J,2006,20(6):711-713.
[8] Rajendran L,Knobloch M,Geiger K D,et al.Increased Abeta production leads to intracellular accumulation of Abeta in flotillin-1-positive endosomes[J].Neurodegener Dis,2007,4(2-3):164-170.
[9] Meister M,Tikkanen R.Endocytic trafficking of membrane-bound cargo:A flotillin point of view[J].Membranes (Basel),2014,4(3):356-371.
[10] Kania E,Pajak B,Gajkowska B,et al.Lipid rafts in Alzheimer's disease[J].Postepy Biochem,2012,58(2):209-216.
[11] Wang X,Yang Q,Guo L,et al.Flotillin-2 is associated with breast cancer progression and poor survival outcomes[J].J Transl Med,2013,11:190.
[12] Wen Q,Wang W,Chu S,et al.Flot-2 expression correlates with EGFR levels and poor prognosis in surgically resected non-small cell lung cancer[J].PLoS One,2015,10(7):e0132190.
[13] Chintagari N R,Gou D,Liu L.Knockdown of flotillin-2 inhibits lung surfactant secretion by alveolar type Ⅱ cells[J]. Cell Res,2008,18(6):701-703.
[14] 刘雨微,成亮,朱剑熹,等.ErbB3和Flotillin-2在骨肉瘤中的表达及临床病理意义[J].中国修复重建外科杂志,2015,5:590-594.
[15] 孙东杰,赵峰,张杰.牛浮舰蛋白-2在真核细胞中的表达与鉴定[J].中国兽医科学,2014,5:463-469.
[16] 朱志,徐惠绵.Flotillin 2能够调节胃癌中HER2水平并与预后相关[A].第9届全国胃癌学术会议暨第二届阳光长城肿瘤学术会议[C].2014.
[17] Wu Y,Zhong A,Zheng H,et al.Expression of flotilin-2 and acrosome biogenesis are regulated by miR-124 during spermatogenesis[J]. PLoS One,2015,10(8):e0136671.
[18] 苏节,刘庆友,朱鹏,等.沼泽型水牛CYP19A1基因克隆、序列分析及组织表达研究[J].畜牧兽医学报,2013,44(4):514-521.
[19] Rajendran L,Masilamani M,Solomon S,et al.Asymmetric localization of flotillins/reggies in preassembled platforms confers inherent polarity to hematopoietic cells[J].Proc Natl Acad Sci USA,2003,100(14):8241-8246.
[20] 谢定芳.Flot-2在胃癌中的表达及其对胃癌细胞增殖和侵袭能力的影响[D].长沙:中南大学,2013.
[21] 张立华.Flotillin-2在鼻咽癌中的表达及作用机制的初步探索[D].长沙:中南大学,2012.
[22] Zhao L,Lin L,Pan C,et al.Flotillin-2 promotes nasopharyngeal carcinoma metastasis and is necessary for the epithelial-mesenchymal transition induced by transforming growth factor-beta[J].Oncotarget,2015,6(12):9781-9793.
[23] 邓廷贤,庞春英,王建,等.水牛分子生物学技术研究新进展[J].中国畜牧兽医,2014,41(4):79-84.
[24] 孙燕,潘红梅,张家骅.水牛胚胎体外生产研究综述[J].中国农学通报,2004,4:17-20.
[25] Possidonio A C,Soares C P,Portilho D M,et al.Differences in the expression and distribution of flotillin-2 in chick,mice and human muscle cells[J].PLoS One,2014,9(8):e103990.
[26] Wu Y,Chen X,Wang S,et al.Flotillin-2 is an acrosome-related protein involved in mouse spermiogenesis[J].J Biomed Res,2012,26(4):278-287.
[27] 陈欣.Flotillin-2在小鼠精子变形过程中的功能研究[D].南京:南京医科大学,2012.

水牛浮舰蛋白2基因克隆及序列分析

Cloning and Sequence Analysis of Buffalo FLOT2 Gene

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	徐媛媛, 郑海英, 杨春艳, 邓廷贤, 黄晨茜, 冯超, 陆杏蓉, 段安琴, 莫霞, 马小娅, 尚江华. 抑制BMP/Smad信号通路对水牛卵巢颗粒细胞生长和类固醇激素合成的影响[J]. 中国畜牧兽医, 2023, 50(6): 2354-2362.
[2]	张方玮, 张琪, 雷亮亮, 孙武胜, 张迪, 张云鹏, 张净搏, 王修权, 张晶, 张树敏. 松辽黑猪HBEGF基因多态性及其与繁殖性状的关联分析[J]. 中国畜牧兽医, 2023, 50(6): 2370-2379.
[3]	肖鹏, 尚江华, 杨春艳, 李孟琪, 段安琴, 马小娅, 冯超, 黄晨茜, 张博, 周金陈, 韦科龙, 郑威, 郑海英. α-亚麻酸对水牛卵巢颗粒细胞体外培养的影响[J]. 中国畜牧兽医, 2023, 50(6): 2380-2387.
[4]	焦琳琳, 成玉婷, 吴庆国, 吴双, 吴植, 朱善元, 钱莺娟. 鸭坦布苏病毒Capsid蛋白原核表达及多克隆抗体制备[J]. 中国畜牧兽医, 2023, 50(6): 2395-2402.
[5]	欧云文, 潘琴, 汪洋, 代军飞, 任绍科, 张洋, 张杰. 猪细小病毒6型ORF2基因的截短表达及多克隆抗体制备[J]. 中国畜牧兽医, 2023, 50(6): 2487-2495.
[6]	刘鹏, 李静静, 乔彦良, 朱连勤, 朱风华. 黄芩多糖的提取及结构表征[J]. 中国畜牧兽医, 2023, 50(6): 2518-2530.
[7]	刘佳隆, 张译文, 张悦然, 孙奇娟, 陈建, 何雷, 贾艳艳, 丁轲, 余祖华. 鸡ELAVL1基因克隆、原核表达及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(5): 1807-1817.
[8]	卓娜, 朱忠武, 李静, 胡世享, 王建昌, 林华, 陈朝林, 韩佃刚, 艾军, 陈培富. 猪源戊型肝炎病毒ORF2蛋白生物信息学分析及真核表达研究[J]. 中国畜牧兽医, 2023, 50(5): 1959-1970.
[9]	王雪平, 张孝俊, 李晓月, 王国栋, 张福良, 宋玉伟, 张明亮, 马磊. 高致病性禽腺病毒血清4型Hexon蛋白的原核表达及多克隆抗体的制备[J]. 中国畜牧兽医, 2023, 50(5): 2053-2060.
[10]	余大新, 刘文静, 刘大鹏, 刘同, 张鹤, 盖凯, 李姝楠, 张贝贝, 侯水生, 周正奎. 鸭胸肌矿物元素含量全基因组关联分析[J]. 中国畜牧兽医, 2023, 50(4): 1287-1295.
[11]	范冰峰, 李文, 刘理想, 赵向远, 邵静, 林乐丰, 孙慧敏, 张旭林, 许保增. 水貂AANAT基因克隆及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(4): 1307-1318.
[12]	刘红润, 朱思燃, 奉玲丽, 张坤, 颜港, 王钰滨, 张帅, 江山, 许迪, 兰干球, 梁晶. 大白猪CDO1基因克隆、生物信息学分析及组织表达定位[J]. 中国畜牧兽医, 2023, 50(4): 1352-1363.
[13]	张蕊, 穆春宇, 汤青萍, 卜柱, 杨明军, 夏爱萍, 杨晓明. 鸽GnIH基因多态性及其与产蛋性状的关联分析[J]. 中国畜牧兽医, 2023, 50(4): 1412-1421.
[14]	张方玮, 张琪, 张云鹏, 李欣, 张净搏, 张树敏, 于永生. 松辽黑猪FOXA2基因第3外显子多态性及其与繁殖性状的关联分析[J]. 中国畜牧兽医, 2023, 50(4): 1444-1451.
[15]	谭元成, 罗华伦, 张依裕, 杨酸, 王盈童, 王春源. 白羽王鸽MyoG基因多态性及其与体尺和肉质性状的关联分析[J]. 中国畜牧兽医, 2023, 50(4): 1461-1471.