[1]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,(04):848-853.[doi:10.3969/j.issn.100-4440.2016.04.021]
　MEI Mei,QIAN Ke,TANG Ying-hua,et al.Fusion expression of subgroup J avian leukosis virus gp85 gene with rabbit IgGFc gene in adenovirus expression system and its application[J].,2016,(04):848-853.[doi:10.3969/j.issn.100-4440.2016.04.021]
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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()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年04期
页码:
848-853
栏目:
畜牧兽医·水产养殖
出版日期:
2016-10-31

文章信息/Info

Title:
Fusion expression of subgroup J avian leukosis virus gp85 gene with rabbit IgGFc gene in adenovirus expression system and its application
作者:
梅梅1; 2; 3; 钱科4; 唐应华1; 3; 秦爱建2; 3; 侯继波1; 3
1. 江苏省农业科学院，江苏 南京 210014；2. 扬州大学江苏省动物预防医学重点实验室，江苏 扬州 225009；3. 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；4. 江苏省出入境检验检疫局，江苏 南京 210001
Author(s):
MEI Mei1; 2; 3; QIAN Ke4; TANG Ying-hua1; 3; QIN Ai-jian2; 3; HOU Ji-bo1; 3
1. Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 2. Key Lab of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; 3. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China; 4. Jiangsu Entry-exit Inspection and Quarantine Bureau, Nanjing 210001, China
关键词:
J亚群禽白血病病毒;  gp85基因; 兔IgGFc; 腺病毒; 融合表达
Keywords:
ubgroup J avian leukosis virus;  gp85 gene;  rabbit rIgGFc;  adenovirus;  fusion expression
分类号:
S858.35.3
DOI:
10.3969/j.issn.100-4440.2016.04.021
文献标志码:
A
摘要:
为获得J亚群禽白血病病毒（ALV-J）囊膜蛋白gp85和兔IgGFc的融合蛋白，通过腺病毒表达系统融合表达ALV-J SUJ和兔IgGFc（rIgGFc）基因。将pcDNA3.1-SUJ-rIgGFc用XhoⅠ、KpnⅠ进行双酶切，获得SUJ-rIgGFc基因，将其克隆至pShuttle-CMV质粒，构建穿梭载体pShuttle-CMV-SUJ-rIgGFc，重组质粒转化含pAdeasy-1的BJ5183-AD-1感受态细胞，获得重组腺病毒质粒pAd-SUJ-rIgGFc，最后转染人293T细胞。表达的融合蛋白可被ALV-J单克隆抗体JE9以及羊抗兔IgG识别，重组蛋白的分子量大小约9.5×10.4，且与JE9及羊抗兔IgG都有很好的反应性；免疫共沉淀试验结果显示融合蛋白可与DF1细胞膜蛋白混合物反应并获得差异蛋白。该融合蛋白可与ALV-J宿主细胞蛋白发生特异性反应。
Abstract:
To obtain the fusion protein of subgroup J avian leukosis virus （ALV-J） envelope protein gp85 and rabbit IgGFc, ALV-J gp85 gene fused with rabbit IgGFc was expressed by adenovirus expression system. The ALV-J gp85 (SU) and rabbit IgGFc (rIgGFc) gene were digested from plasmid pcDNA3.1-SUJ-rIgGFc with XhoⅠand KpnⅠand cloned into pShuttle-CMV to construct the recombinant plasmid pShuttle-CMV-SUJ-rIgGFc which was transformed into BJ5183-AD-1 competent cells containing pAdeasy-1 to construct the recombinant adenovirus plasmid pAd-SUJ-rIgGFc. The recombinant adenovirus was obtained by transfecting the recombinant plasmid pAd-SUJ-rIgGFc into 293T cells. The fusion protein can be recognized by mAb JE9 specific to gp85 of ALV-J and antibodies against rabbit IgG with the molecular weight about 9.5×10.4. The result of Co-IP showed that the fusion protein SUJ-rIgGFc reacted with the membrane proteins from DF1 cells and produced differential proteins, indicative of the specific reaction between fused protein and the protein from host cells of ALV-J.

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备注/Memo

备注/Memo:
收稿日期：2015-12-14基金项目：国家自然科学基金青年基金项目（31201881）；教育部创新团队项目（IRT0978）；江苏省普通高校研究生科研创新计划项目（CXL2_0938X1）作者简介：梅梅（1982-），女，江苏东台人，博士研究生，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: 2016-11-01