Expression and Genic Imm unization of Glycoprotein gB Gene ofM arek’s disease virus

DING Qiao; LI Jian-jun; CHEN Pu

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June 2003

DING Qiao, LI Jian-jun and CHEN Pu. Expression and Genic Imm unization of Glycoprotein gB Gene ofM arek’s disease virus[J]. Virologica Sinica, 2003, 18(3): 275-278.

Citation: DING Qiao, LI Jian-jun, CHEN Pu. Expression and Genic Imm unization of Glycoprotein gB Gene ofM arek’s disease virus .VIROLOGICA SINICA, 2003, 18(3) : 275-278.

鸡马立克氏病病毒糖蛋白gB基因的表达及基因免疫

丁巧玲 ¹ ,
李建军 ² ,
陈溥言 ^1*

1.
1．南京农业大学农业部动物疫病诊断与免疫重点开发实验室，江苏南京，210095
2.
佛山科技学院动物医学系，广东南海，528231

摘要

将马立克氏病病毒(Marek’s disease virus,MDV)gB基因插入真核表达载体pcDNA3中,经酶切和PCR鉴定为正向插入了gB基因的重组质粒,命名为pcDNA3-gB。体外转染COS-7细胞,经间接免疫荧光染色证实转染pcDNA3-gB后的COS-7细胞内有糖蛋白B的表达。用pcDNA3-gB对AA肉用雏鸡腿部肌肉注射,进行一免。间隔2周后,用pcDNA3-gB再加强免疫一次。之后2周攻毒,观察免疫保护效果。结果表明,MDV gB基因DNA免疫可以诱导鸡体产生免疫保护,免疫保护指数为72.2%。
- 马立克氏病病毒（Marek\’s
- , disease
- , virus
- , MDV）
- , gB基因
- , 表达
- , 基因免疫

Expression and Genic Imm unization of Glycoprotein gB Gene ofM arek’s disease virus

DING Qiao ¹ ,
LI Jian-jun ² ,
CHEN Pu ^1*

1.
Key Laboratory of Animal Diseases Diagnostic and Immunology of Agriculture Ministry of China,Nanjing Agricultural University,Nanjing 210095．China
2.
Department of Veterinary medicine,Foshan Science＆Technology College，Nanhai 528231，Chinaf1．Key Laboratory of Animal Diseases Diagnostic and Immunology of Agriculture Ministry of China,Nanjing Agricultural University,Nanjing 210095．China

Abstract

gB gene of Marek’s disease virus (MDV) was inserted into pcDNA3, forming eukaryotic expression recombinant plasmid pcDNA3-gB. Recombinant plasmid pcDNA3-gB was identified by BamHⅠI and PCR. PcDNA3-gB was transfected into COS-7 cells in vitro. Expressed protein was detected by indirect immunofluorescence assay. Immune test groups were inoculated with pcDNA3-gB intramuscularly. Two weeks later, test groups were boosted with the same pcDNA3-gB. Since then, two weeks later chickens were challenged with vMDV. Immune effects showed that the immune protection rate of test group reached to 72.2%. The results proved that pcDNA3-gB DNA immunization elicited immune protection.
- Marek\’s disease virus（MDV） gB gene Expression DNA vaccine

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression and Genic Imm unization of Glycoprotein gB Gene ofM arek’s disease virus

1. Key Laboratory of Animal Diseases Diagnostic and Immunology of Agriculture Ministry of China,Nanjing Agricultural University,Nanjing 210095．China
2. Department of Veterinary medicine,Foshan Science＆Technology College，Nanhai 528231，Chinaf1．Key Laboratory of Animal Diseases Diagnostic and Immunology of Agriculture Ministry of China,Nanjing Agricultural University,Nanjing 210095．China

Abstract: gB gene of Marek’s disease virus (MDV) was inserted into pcDNA3, forming eukaryotic expression recombinant plasmid pcDNA3-gB. Recombinant plasmid pcDNA3-gB was identified by BamHⅠI and PCR. PcDNA3-gB was transfected into COS-7 cells in vitro. Expressed protein was detected by indirect immunofluorescence assay. Immune test groups were inoculated with pcDNA3-gB intramuscularly. Two weeks later, test groups were boosted with the same pcDNA3-gB. Since then, two weeks later chickens were challenged with vMDV. Immune effects showed that the immune protection rate of test group reached to 72.2%. The results proved that pcDNA3-gB DNA immunization elicited immune protection.