PRRSV 2b蛋白与LGALS1相互作用的验证研究

doi:10.11924/j.issn.1000-6850.casb17090090

中国农学通报 ›› 2018, Vol. 34 ›› Issue (29): 112-117.doi: 10.11924/j.issn.1000-6850.casb17090090

所属专题：生物技术

PRRSV 2b蛋白与LGALS1相互作用的验证研究

杨达汉,邓守全,崔一龙,刘锴,王学理,霍晓伟,马德慧,薛江东

内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院,内蒙古民族大学动物科学技术学院

收稿日期:2017-09-15 修回日期:2017-12-19 接受日期:2018-01-24 出版日期:2018-10-15 发布日期:2018-10-15
通讯作者: 马德慧
基金资助:
国家自然科学基金项目“猪繁殖与呼吸综合征病毒2b 蛋白与宿主细胞蛋白相互作用研究”(31460662)；内蒙古自治区自然科学基金项目 “2b 蛋白介导猪繁殖与呼吸综合征病毒感染易感细胞的分子机制研究”(2017MS0319)；内蒙古自治区高等学校科学研究项目“猪繁殖与呼吸综合征病毒2b 蛋白介导的宿主细胞离子通道变化研究”(NJZY13163)。

The Validation Research on Interaction Between PRRSV 2b Protein and LGALS1

Received:2017-09-15 Revised:2017-12-19 Accepted:2018-01-24 Online:2018-10-15 Published:2018-10-15

摘要/Abstract

摘要： 旨在研究猪繁殖与呼吸综合症病毒(Porcine reproductive and respiratory syndrome virus，PRRSV)2b 蛋白与宿主细胞蛋白之间的相互作用。本研究利用酵母回返试验、GST-pull down 试验、免疫共沉淀试验3 种方法对LGALS1 与PRRSV 2b 蛋白间是否相互作用进行验证。结果表明：在酵母回返试验中，含有LGALS1 与2b 基因的杂交酵母菌株在SD/-Trp-Leu-His-Ade/AbA/X-α-gal 的平板上生长出蓝色菌落；在GST-pull down 试验中，LGALS1 与2b 蛋白反应的样品经Western blot 可检测到融合蛋白带；在免疫共沉淀试验中，LGALS1 与2b 蛋白反应的样品经HA单抗沉淀后，经Western blot 可检测到相应蛋白带。该研究证明了LGALS1 与PRRSV 2b 蛋白间发生相互作用，为研究PRRSV感染途径和增殖过程提供了理论基础。

关键词: 甜菜, 甜菜, 杂种优势, 遗传机理, 遗传距离, 育种

Abstract: The aim is to study the interaction between porcine reproductive and respiratory syndrome virus (PRRSV) 2b protein and host cell proteins. In this study, we used yeast regression test, GST-pull down assay and co- immunoprecipitation assay to verify the interaction between LGALS1 and PRRSV 2b. The results showed that in the yeast regression test, hybrid yeast strain of LGALS1 and2b gene grew blue yeast colonies on SD/-Trp-Leu-His-Ade/AbA/X-α-gal plates. In the GST-pull down assay, the fusion protein bands were detected by Western blot in the samples reacted with 2b protein and LGALS1. In the co-immunoprecipitation assay, samples reacted with 2b protein and LGALS1 were precipitated with HA monoclonal antibody and the corresponding protein bands were detected by Western blot. This paper demonstrates the interaction between LGALS1 and PRRSV 2b protein and provides a theoretical basis for studying PRRSV infection and proliferation.

杨达汉,邓守全,崔一龙,刘锴,王学理,霍晓伟,马德慧,薛江东. PRRSV 2b蛋白与LGALS1相互作用的验证研究[J]. 中国农学通报, 2018, 34(29): 112-117.

参考文献

1 刘莹,王凤雪,温永俊,等. 猪繁殖与呼吸综合征病毒非结构蛋白2的研究进展[J]. 中国预防兽医学报,2015,(02):155-158.
2 田志平,薛江东,马徳慧. 猪繁殖与呼吸综合征病毒E蛋白研究进展[J]. 中国畜牧兽医,2012,39(10):42-44.
3 Wang Q, Li Y, Dong H, et al. Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain[J]. Virol J. 2017,14(1):356-272.
4 Zhang K, Hou Q, Zhong Z, et al. Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E[J]. Virology. 2013, 442(2):156-162.
5 Zhou Y, Fang L, Wang D, et al. Porcine Reproductive and Respiratory Syndrome Virus Infection Induces Stress Granule Formation Depending on Protein Kinase R-like Endoplasmic Reticulum Kinase(PERK) in MARC-145 Cells[J]. Front Cell Infect Microbiol. 2017,4(7):31-37.
6 Martínez-Bosch N, Fernández-Barrena MG, Moreno M, et al.Galectin-1 drives pancreatic carcinogenesis through stroma remodeling and Hedgehog signaling activation[J]. Cancer Res. 2014, 74(13):3512-3524.
7 Park GB, Chung YH, Kim D. Induction of galectin-1 by TLR-dependent PI3K activation enhances epithelial-mesenchymal transition of metastatic ovarian cancer cells[J]. Oncol Rep. 2017, 37(5):3137-3145.
8 Thiemann S, Baum LG. Galectins and Immune Responses-Just How Do They Do Those Things They Do?[J]. Annu Rev Immunol. 2016, 34:243-264.
9杨达汉,邓守全,王哲,等. 猪肺泡巨噬细胞酵母双杂交cDNA文库的构建与鉴定[J]. 中国兽医学报,2017,37(04):632-636+642.
10 ]李先昆,聂智毅,曾日中. 酵母双杂交技术研究与应用进展[J]. 安徽农业科学,2009,37(07):2867-2869+2926.
11 Martínez-Bosch N, Fernández-Barrena MG, Moreno M, et al.Galectin-1 drives pancreatic carcinogenesis through stroma remodeling and Hedgehog signaling activation[J]. Cancer Res. 2014, 74(13):3512-3524.
12 Valach J, Fik Z Strnad H, et al. Smooth muscle actin-expressing stromal fibroblasts in head and neck squamous cell carcinoma: Increased expression of galectin-1 and induction of poor prognosis factors[J]. Int. J. Cancer. 2012, 131, 2499–2508.
13 Yang N, Zhang W, He T, et al. Suppression of Retinal Neovascularization by Inhibition of Galectin-1 in a Murine Model of Oxygen-Induced Retinopathy[J]. J Ophthalmol. 2017, 119(5):224-232.
14 Huang YJ, Shiau AL, Chen SY, et al. Multivalent structure of galectin-1-nanogold complex serves as potential therapeutics for rheumatoid arthritis by enhancing receptor clustering[J]. Eur. Cells Mater. 2012, 23, 170–181.
15 D’Haene N, Sauvage S, Maris C, et al. VEGFR1 and VEGFR2 involvement in extracellular galectin-1- and galectin-3-induced angiogenesis. PLoS ONE, 2013, 8, e67029.
16 Park GB, Chung YH, Kim D. Induction of galectin-1 by TLR-dependent PI3K activation enhances epithelial-mesenchymal transition of metastatic ovarian cancer cells[J]. Oncol Rep. 2017, 37(5):3137-3145.
17 Nam K, Son SH, Oh S, et al. Binding of galectin-1 to integrin β1 potentiates drug resistance by promoting survivin expression in breast cancer cells[J]. Oncotarget. 2017, 92, 1188–1192.
18 Ose R, Oharaa O, Nagase T. Galectin-1 and galectin-3 mediate protocadherin-24-dependent membrane localization of catenin in colon cancer cell line HCT116[J]. Curr. Chem. Genom. 2012, 6, 18–26.
19 Astorgues-Xerri L, Riveiro ME, Tijeras-Raballand,et al. Unraveling galectin-1 as a novel therapeutic target for cancer[J]. Cancer Treat. Rev. 2014, 40, 307–319.
20 Geiger P, Mayer B, Wiest I,et al. Binding of galectin-1 to breast cancer cells MCF7 induces apoptosis and inhibition of proliferation in vitro in a 2D- and 3D- cell culture model[J]. BMC Cancer. 2016, 16(1):870.
21 Takahashi H(1), Takahashi C, Moreland NJ, et al. Establishment of a robust dengue virus NS3-NS5 binding assay for identification of protein-protein interaction inhibitors[J]. Antiviral Res. 2012, 96(3):305-314.
22 Soellick, T.R, Uhrig J.F. Development of an optimized interaction-mating protocol for large-scale yeast two-hybrid analyses[J]. Genome Biology. 2011, 7(4): 1-18.
24 Fong BA, Gillies AR, Ghazi I, et al. Purification of Escherichia coli RNA polymerase using a self-cleaving elastin-like polypeptide tag[J]. Protein Sci. 2010, 19(6):1243-52.

[1]	贾也纯, 陈润仪, 贺泽霖, 倪洪涛. 甜菜抗非生物胁迫研究进展[J]. 中国农学通报, 2022, 38(9): 33-40.
[2]	陈英花, 白如霄, 王娟, 张新疆, 刘玲慧, 刘小龙, 冯国瑞, 危常州. 叶面喷施烯效唑和硼对塔额盆地甜菜产量和含糖率的影响[J]. 中国农学通报, 2022, 38(9): 41-48.
[3]	巩永永, 端木慧子. 甜菜TIFY基因家族的全基因组鉴定与生物信息学分析[J]. 中国农学通报, 2022, 38(8): 17-24.
[4]	王琳玉, 蒋依辰, 于清洋, 吴则东, 邳植. 甜菜组蛋白去乙酰化酶(HDACs)基因家族鉴定及功能预测[J]. 中国农学通报, 2022, 38(8): 9-16.
[5]	邓裕帅, 王宇光, 於丽华, 耿贵. 水涝胁迫对不同土壤盐碱度下甜菜幼苗生长及光合特性的影响[J]. 中国农学通报, 2022, 38(7): 18-23.
[6]	姚洁, 程磊, 周涛, 雷鹏堃, 朱月健, 马磊. 国内太子参种质资源与遗传育种研究进展[J]. 中国农学通报, 2022, 38(7): 62-66.
[7]	梁增基, 慕芳, 王楠. 渭北高原小麦育种的演变与展望[J]. 中国农学通报, 2022, 38(6): 8-14.
[8]	闫蕴韬, 何兮, 张海清, 贺记外. 水稻种子耐贮性研究进展[J]. 中国农学通报, 2022, 38(5): 1-8.
[9]	王盛昊, 于冰. 甜菜M14品系BvM14-UNG基因克隆及生物信息学分析[J]. 中国农学通报, 2022, 38(4): 16-22.
[10]	刘镎, 胡华兵, 王荣华, 刘小越, 刘朝阳, 刘晓晗, 王茂芊. 甲醇老化处理对甜菜种子发芽的影响[J]. 中国农学通报, 2022, 38(33): 28-33.
[11]	王佳琦, 张子萱, 刘乃新. 外源硒处理条件下红甜菜苗期矿物质积累特性分析[J]. 中国农学通报, 2022, 38(32): 1-5.
[12]	赵雅儒, 邳植, 刘蕊, 马语嫣, 吴则东. 不同甜菜单胚细胞质雄性不育系与保持系的遗传多样性分析[J]. 中国农学通报, 2022, 38(30): 35-40.
[13]	董寅壮, 王堽, 於丽华, 耿贵. 亚铁胁迫对甜菜幼苗矿质元素积累的影响[J]. 中国农学通报, 2022, 38(3): 11-16.
[14]	石杨, 尹希龙, 李王胜, 兴旺. PEG模拟干旱胁迫对耐旱型与干旱敏感型甜菜种质形态指标的影响[J]. 中国农学通报, 2022, 38(29): 45-51.
[15]	唐玲, 孙思琦, 闫桦, 刘亚昕, 刘大丽. 甜菜红素提取与纯化技术的研究进展[J]. 中国农学通报, 2022, 38(28): 136-142.

PRRSV 2b蛋白与LGALS1相互作用的验证研究

The Validation Research on Interaction Between PRRSV 2b Protein and LGALS1

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

关于本刊

作者中心

审者中心

在线期刊

联系我们