[1]古努尔·吐尔逊,王登峰,杨学云,等.梅迪-维斯纳病毒gag蛋白的原核表达及其交叉反应原性[J].江苏农业学报,2020,(04):978-983.[doi:doi:10.3969/j.issn.1000-4440.2020.04.024]
　GUNUER Tuerxun,WANG Deng-feng,YANG Xue-yun,et al.Prokaryotic expression and cross reactogenicity of maedi-visna virus gag protein[J].,2020,(04):978-983.[doi:doi:10.3969/j.issn.1000-4440.2020.04.024]
点击复制

梅迪-维斯纳病毒gag蛋白的原核表达及其交叉反应原性()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年04期
页码:
978-983
栏目:
畜牧兽医·水产养殖
出版日期:
2020-08-31

文章信息/Info

Title:
Prokaryotic expression and cross reactogenicity of maedi-visna virus gag protein
作者:
古努尔·吐尔逊; 王登峰; 杨学云; 魏玉荣; 李建军; 孟肖潇; 洪都孜·波拉提; 吴建勇
(新疆畜牧科学院兽医研究所，新疆乌鲁木齐830013）
Author(s):
GUNUER Tuerxun; WANG Deng-feng; YANG Xue-yun; WEI Yu-rong; LI Jian-jun; MENG Xiao-xiao; HONGDUZI Bolati; WU Jian-yong
(Institute of Veterinary Medicine, Xinjiang Academy of Animal Sciences, Urumqi 830013, China）
关键词:
梅迪-维斯纳病毒; gag蛋白; 原核表达; 交叉反应原性; 抗原表位
Keywords:
maedi-visna virus; gag protein; prokaryotic expression; cross reactogenicity; epitope
分类号:
S852.65
DOI:
doi:10.3969/j.issn.1000-4440.2020.04.024
文献标志码:
A
摘要:
为鉴定适用于中国小反刍动物慢病毒(Small ruminants lentivirus viruses，SRLVs）血清学诊断的通用蛋白标记物，本研究以梅迪-维斯纳病毒(Maedi-Visna virus，MVV）四川株为模板扩增获得gag基因序列，使用生物信息学方法分析其与中国山羊关节炎-脑炎病毒(Caprine arthritis encephalitis virus，CAEV）分离株gag蛋白氨基酸序列和抗原表位相似性，构建gag基因重组表达载体并转入BL21(DE3)宿主菌进行诱导表达，采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE）和Western blotting确定其大小、分布和反应原性。结果显示，MVV gag蛋白氨基酸序列与中国CAEV分离株的相似性为74.7%～74.9%，它的11个抗原表位与CAEV gag蛋白的12个抗原表位存在较高的序列相似性，推测其可作为中国SRLVs血清学诊断的通用蛋白质标记物；SDS-PAGE检测结果显示，重组gag蛋白大小约为55 000，主要以包涵体形式存在，Western blotting显示该蛋白可与MVV和CAEV感染动物血清发生免疫反应，证实了蛋白序列分析结果。该研究结果可为研制中国SRLVs通用血清学检测技术方法奠定基础。
Abstract:
To identify the universal protein biomarker for serological diagnosis of small ruminant lentivirus (SRLVs) in China, the gag gene was amplified from maedi-visna virus (MVV) Sichuan strain, and the similarity of its amino acid sequence and epitopes between MVV Sichuan strain and caprine arthritis encephalitis virus (CAEV) strain in China was analyzed by bioinformatics method. The recombinant expression vector of gag gene was then constructed and transferred into Escherichia Coli BL21 (DE3), SDS-PAGE and western blotting were used to determine the size, distribution and reactogenicity of recombinant gag protein. The results showed that the amino acid sequence of MVV gag protein was 74.7%-74.9% similar to that of CAEV strain in China, and its 11 epitopes were highly similar to 12 epitopes of CAEV gag protein, suggesting that MVV gag protein could be used as the universal protein biomarker for serodiagnosis of SRLVs in China. The results of SDS-PAGE detection indicated that the recombinant gag protein was about 55 000 in size, mainly in the form of inclusion body. Western blotting detection results showed that the protein could react with the sera of MVV and CAEV infected animals, which verified the results of protein sequence analysis. These results lay a foundation for the development of a universal serological detection method for SRLVs in China.

参考文献/References:

[1]易海清，阿合买提·买买提，邓普辉，等. 新疆卡拉库尔羊梅迪/维斯那病毒感染的病理学观察[J]. 动物医学进展，2006，27(2）：75-78.
[2]SIGURDSSON B, THORMAR H, PLSSON P A. Cultivation of Visna virus in tissue culture[J]. Archives of virology, 1960, 10(3）:368-381.
[3]GOMEZ-LUCIA E, BARQUERO N, DOMENECH A. Maedi-Visna virus: current perspectives[J]. Veterinary Medicine-Research and Reports, 2018, 9:11-21.
[4]ARNARSON H, PALSSON A, GUDNADOTTIR M, et al. Maedi-Visna virus persistence: antigenic variation and latency[J]. Comparative Immunology Microbiology and Infectious Diseases, 2017, 55: 6-12.
[5]OGUMA K, TANAKA C, HARASAWA R, et al. Isolation of Maedi/Visna virus from a sheep in Japan[J]. Journal of Veterinary Medical Science, 2014, 76(2): 211-218.
[6]LAGO N, LOPEZ C, PANADERO R, et al. Seroprevalence and risk factors associated with Visna/Maedi virus in semi-intensive lamb-producing flocks in northwestern Spain[J]. Preventive Veterinary Medicine, 2012, 103(2/3): 163-169.
[7]PEREZ M, BIESCAS E, DE ANDRES X, et al. Visna/maedi virus serology in sheep Survey, risk factors and implementation of a successful control programme in Aragon (Spain) [J]. Veterinary Journal, 2010, 186(2): 221-225.
[8]ALBAYRAK H, YAZICI Z, OKUR-GUMUSOVA S, et al. Maedi-visna virus infection in Karayaka and Amasya Herik breed sheep from provinces in northern Turkey[J]. Tropical Animal Health and Production, 2012, 44(5): 939-941.
[9]AZKUR A K, GAZYAGCI S, ASLAN M E. Serological and epidemiological investigation of bluetongue, Maedi-Visna and caprine arthritis-encephalitis viruses in small ruminant in kirikkale district in Turkey[J]. Kafkas Universitesi Veteriner Fakultesi Dergisi, 2011, 17(5): 803-808.
[10]PREZIUSO S, OR M E, GIAMMARIOLI M, et al. Maedi-visna virus in Turkish sheep: a preliminary serological survey using ELISA tests[J]. Turkish Journal of Veterinary & Animal Sciences, 2010, 34(3): 289-293.
[11]陈爱素，沙音，董宝山. 从绵羊进行性肺炎病例中分离提纯梅迪(Maedi)病毒及其形态观察[J]. 中国兽医杂志，1984，10(2）：4-5.
[12]王进成，符子华，胡尔玛西，等. 绵羊进行性肺炎流行的血清学调查[J]. 中国兽医杂志，1997，23(4）：28-29.
[13]孙雨，杨林，王传彬，等. 我国11个省份山羊关节炎-脑炎与绵羊进行性肺炎的血清抗体检测[J]. 中国兽医科学，2018，48(1)：34-38.
[14]WORKMAN A M, DICKEY A M, HEATON M P, et al. Complete genome sequences of two genotype A2 small ruminant lentiviruses isolated from infected U.S. sheep[J]. Microbiology Resource Announcements, 2017, 5(13): e00109.
[15]RAMIREZ H, REINA R, AMORENA B, et al. Small ruminant lentiviruses: genetic variability, tropism and diagnosis[J]. Viruses-Basel, 2013, 5(4): 1175-1207.
[16]HIGHLAND M A. Small ruminant lentiviruses: strain variation, viral tropism, and host genetics influence pathogenesis[J]. Veterinary Pathology, 2017, 54(3): 353-354.
[17]邓普辉，王正党，苏忠，等. 中国绵羊进行性肺炎(梅迪)流行的血清学调查[J]. 中国兽医杂志，1989，15(4）：2-4.
[18]RECHE P A. Predicted Antigenic Peptides [DB/OL]. (2002-06-01）
[2020-02-03]. http://imedmeducmes/Tools/antigenicpl.
[19]杨义琴，吴建勇，古努尔·吐尔逊，等. 小反刍动物慢病毒通用PCR检测方法的建立与初步评价[J]. 草食家畜，2018(2）：13-19.
[20]林杰，胡泽渊，胡尔玛西，等. 山羊关节炎脑炎的血清学诊断[J]. 中国兽医杂志，1987， 13(10）：12-14.
[21]胡尔马希，林杰，胡泽渊，等. 山羊关节炎脑炎病毒的分离与鉴定[J]. 中国兽医科技，1989(10）：5-6.
[22]陈欢欢. 聚乙二醇沉淀法在原核表达系统生物技术药物蛋白分离中的应用研究[D]. 上海：上海交通大学，2017.

备注/Memo

备注/Memo:
收稿日期：2020-02-03基金项目：国家重点研发项目(2016YF0500908）作者简介：古努尔·吐尔逊(1971-），女，维吾尔族，新疆伊犁人，学士，高级实验师，主要从事动物传染病研究。(E-mail）azn120219@126.com通讯作者：吴建勇，(E-mail）chienyung@foxmail.com
更新日期/Last Update: 2020-09-08