[1]梅梅,胡晓田,秦爱建,等.A亚群禽白血病病毒env基因在昆虫细胞中的表达与鉴定[J].江苏农业学报,2017,(06):1316-1320.[doi:doi:10.3969/j.issn.1000-4440.2017.06.017]
　MEI Mei,HU Xiao-tian,QIN Ai -jian,et al.Expression and identification of subgroup A avian leukosis virus env in insect cell[J].,2017,(06):1316-1320.[doi:doi:10.3969/j.issn.1000-4440.2017.06.017]
点击复制

A亚群禽白血病病毒env基因在昆虫细胞中的表达与鉴定()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年06期
页码:
1316-1320
栏目:
畜牧兽医·水产养殖
出版日期:
2017-12-30

文章信息/Info

Title:
Expression and identification of subgroup A avian leukosis virus env in insect cell
作者:
梅梅1; 2; 胡晓田2; 3; 秦爱建2; 3; 陆吉虎1; 2; 张雪花1; 2; 侯继波1; 2; 唐应华1; 2
（1.江苏省农业科学院动物免疫工程研究所，江苏南京210014；2.江苏省动物重要疫病与人兽共患病防控协同创新中心，江苏扬州225009；3.扬州大学江苏省动物预防医学重点实验室，江苏扬州225009）
Author(s):
MEI Mei1; 2; HU Xiao-tian2; 2; QIN Ai -jian2; 3; LU Ji-hu1; 2; ZHANG Xue-hua1; 2; HOU Ji-bo1; 2; TANG Ying-hua1; 2
(1.Institute of Veterinary Immunology and Engineering， Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;3.Key lab of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou 225009, China)
关键词:
A亚群禽白血病病毒; env基因; 表达
Keywords:
avian leukosis virus subgroup A（ALV-A）; env gene; expression
分类号:
S855.3
DOI:
doi:10.3969/j.issn.1000-4440.2017.06.017
文献标志码:
A
摘要:
为获得A亚群禽白血病病毒（ALV-A）env基因的表达产物，以ALV-A AH10毒株病毒RNA为模板，通过反转录PCR扩增得到env基因，将其克隆到转移载体pFastBac1中，转化DH10Bac感受态细胞，获得重组穿梭质粒rBacmid-envA，rBacmid-envA转染昆虫细胞Sf9，获得重组杆状病毒。间接免疫荧光试验（IFA）和免疫印迹试验（Western-blot）结果显示，重组蛋白可与抗ALV-A抗体发生特异性反应，重组蛋白分子大小约为90 000，与预期大小相符，表明env基因在Sf9细胞中获得了良好表达。
Abstract:
To abtain the expression product of subgroup A avian leukosis virus env gene, the env gene amplified from subgroup A avian leukosis virus strain AH10 was cloned into transfer vector pFastBac1 and cloned into the shuttle vector Bacmid by transferred into DH10Bac competent cell. The recombinant shuttle plasmid rBacmid-envA was then transfected into Sf9 cells to generate recombinant virus. The results of indirect immunofluorescence assay (IFA) and western blot showed that the recombinant protein could be recognized specifically by the polyclonal antibody against ALV-A with the molecule weight about 90 000, which was consistent with the expected, and all the results indicated that the env gene was expressed well in Sf9 cells .

参考文献/References:

[1]SHAO H X, WANG L, SANG J J, et al. Novel avian leukosis viruses from domestic chicken breeds in mainland China[J]. Archives of Virology, 2017, 162(7)：2073-2076.
[2]孙鹏，陈孜孟，赵国梁，等. K亚群禽白血病病毒gp85单因子血清的制备及其特异性鉴定[J]. 中国畜牧兽医，2016，43（2）：371-376.
[3]塞弗. 禽病学 [M]. 11版. 北京：中国农业出版社， 2005.
[4]CUI N, SU S, CHEN Z M, et al. Genomic sequence analysis and biological characteristics of a rescued clone of avian leukosis virus strain JS11C1, isolated from indigenous chickens[J]. Journal of General Virology, 2014, 95:2512-2522.
[5]LI X, LIN W, CHANG S, et al. Isolation, identification and evolution analysis of a novel subgroup of avian leukosis virus isolated from a local Chinese yellow broiler in South China[J]. Archives of Virology, 2016, 161(10): 2717-2725.
[6]ONO M, TSUKAMOTO K, TANIMURA N, et al. An epizootic of subcutaneous tumors associated with subgroup A avian leukosis/sarcoma virus in young layer chickens[J]. Avian Diseases, 2004, 48(4): 940-946.
[7]ZHANG Q C, ZHAO D M, GUO H J, et al. Isolation and identification of a subgroup A avian leukosis virus from imported meat-type grand-parent chickens[J]. Virologica Sinica, 2010, 25(2): 130-136.
[8]王培坤，毕玉彧，秦丽莉，等. 三黄鸡临床血管瘤病例中分离出A亚群与J亚群禽白血病病毒[J]. 动物医学进展，2015，36（3）：13-16.
[9]冯敏，谭利强，代曼曼，等. 种禽场A亚群禽白血病病原学调查及分离株遗传进化分析[J]. 华南农业大学学报，2014，35（4）：11-15.
[10]张恒，李传龙，杨明，等.禽白血病A亚群gp85的单因子血清制备及其特异性鉴定[J].微生物学报，2011，51（1）：134-140.
[11]钱琨，朱钰峰，沈海钰，等. 地方蛋鸡群中A亚群禽白血病病毒的分离与全基因组序列分析[J]. 中国兽医科学，2011，41（10）：1005-1010.
[12]FENG M, DAI M M, LIAO M, et al. Establishment of an avian leukosis virus subgroup A-resistant cell line[J]. Journal of Integrative Agriculture, 2017, 16(4):930-936.
[13]PAYNE L N, NAIR V. The long view: 40 years of avian leukosis research[J]. Avian Pathology, 2012, 41(1): 11-19.
[14]BOVA C A, OLSEN J C, SWANSTROM R. The avian retrovirus env gene family: molecular analysis of host range and antigenic variants[J]. Journal of Virology, 1988, 62(1): 75-83.
[15]魏彦辉，周荣艳，李祥龙，等. 抗A亚群禽白血病受体基因在三个鸡群中分布的研究[J]. 黑龙江畜牧兽医，2017（3）：167-170.
[16]梅梅，钱科，唐应华，等. J亚群禽白血病病毒SUJ及兔IgGFc基因在腺病毒表达系统中的融合表达［ Ｊ］.江苏农业学报， 2016，32（4）：848-853.
[17]SUN S, CUI Z. Epidemiological and pathological studies of subgroup J avian leukosis virus infections in Chinese local ‘yellow’ chickens[J]. Avian Pathology, 2007, 36(3): 221-226.
[18]ZAVALA G, CHENG S. Experimental infection with avian leukosis virus isolated from Marek′s disease vaccines[J]. Avian Diseases, 2006, 50(2): 232-237.

备注/Memo

备注/Memo:
收稿日期：2017-07-12 基金项目：国家自然科学基金项目（31201881） 作者简介：梅梅（1982-），女，江苏东台人，博士，助理研究员，主要从事动物免疫学研究。（E-mail）jacqui18@163.com 通讯作者：秦爱建，（E-mail）aijian@yzu.edu.cn；唐应华，（E-mail）tyinghua@126.com
更新日期/Last Update: 2018-01-03