Construction of Recombinant Marek’s Disease Virus Vaccine Strain CVI988/Rispens Expressing the VP2 Protein of IBDV

LIU Hong-mei; QIN Ai-jian*; YE Jian-qiang; CHEN Hong-jun; SHAO Hong-xia; JIN Wen-jie; LIU Yue-long

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June 2006

LIU Hong-mei, QIN Ai-jian*, YE Jian-qiang, CHEN Hong-jun, SHAO Hong-xia, JIN Wen-jie and LIU Yue-long. Construction of Recombinant Marek’s Disease Virus Vaccine Strain CVI988/Rispens Expressing the VP2 Protein of IBDV[J]. Virologica Sinica, 2006, 21(3): 257-260.

Citation: LIU Hong-mei, QIN Ai-jian*, YE Jian-qiang, CHEN Hong-jun, SHAO Hong-xia, JIN Wen-jie, LIU Yue-long. Construction of Recombinant Marek’s Disease Virus Vaccine Strain CVI988/Rispens Expressing the VP2 Protein of IBDV .VIROLOGICA SINICA, 2006, 21(3) : 257-260.

表达传染性法氏囊病毒VP2蛋白的重组马立克氏病毒的构建

1.
扬州大学江苏省动物预防医学重点实验室，江苏扬州 225009

摘要

用聚合酶链式反应（PCR）方法从真核表达载体pcDNA3.1-VP2中扩增出包含CMV和polyA的VP2表达盒基因片段，经琼脂糖凝胶电泳大小为2.4kb；将该基因片段插入马立克氏病病毒（MDV）CVI988//Rispens的非必需区US10片段中，经SphI酶切分析获得大小为6.0和2.4kb两个片段，表明成功构建出表达VP2基因的MDV CVI988转移载体pUC18-US10-VP2质粒。将质粒与CVI988/Rispens疫苗毒共转染鸡胚成纤维细胞（CEF），通过免疫荧光方法筛选，结果获得了表达VP2基因的重组MDV（rMDV-VP2）。用IBDV特异性单克隆抗体进行间接免疫荧光试验（IFA）证实，rMDV-VP2病毒传至第8代仍能稳定表达VP2蛋白,这为进一步研究其免疫学特性奠定了基础。
- 马立克氏病毒（Marek’s disease virus, MDV）
- , 转移载体
- , 重组病毒
- , VP2

Construction of Recombinant Marek’s Disease Virus Vaccine Strain CVI988/Rispens Expressing the VP2 Protein of IBDV

1.
Key Lab of Jiangsu preventive veterinary medicine, Yangzhou University, Yangzhou 225009, China

Abstract

An expression cassette containing the VP2 gene of infectious bursal disease virus (IBDV) strain JS under the control of the CMV promoter and an enhancer in pcDNA3.1-VP2 was amplified by PCR and cloned into pUC18-US10 plasmid to yield the transferring vector pUC18-US10-VP2. The recombinant virus, designated rCVI988-VP2, was generated by co-transfection of CEF with pUC18-US10-VP2 and the Marek’s disease virus (MDV) vaccine CVI988/Rispens virus. PCR analysis and immunofluorenscence assays indicated that the recombinant virus was stable over 8 passages and that VP2 was expressed in CEF infected with the rCVI988-VP2. The data showed that the recombinant MDV expressing the VP2 gene of IBDV was successfully constructed and could prove to be a good strategy for IBD control.
- Marek’s disease virus
- , VP2
- , Transferring vector
- , Recombinant virus

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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

Construction of Recombinant Marek’s Disease Virus Vaccine Strain CVI988/Rispens Expressing the VP2 Protein of IBDV

1. Key Lab of Jiangsu preventive veterinary medicine, Yangzhou University, Yangzhou 225009, China

Abstract: An expression cassette containing the VP2 gene of infectious bursal disease virus (IBDV) strain JS under the control of the CMV promoter and an enhancer in pcDNA3.1-VP2 was amplified by PCR and cloned into pUC18-US10 plasmid to yield the transferring vector pUC18-US10-VP2. The recombinant virus, designated rCVI988-VP2, was generated by co-transfection of CEF with pUC18-US10-VP2 and the Marek’s disease virus (MDV) vaccine CVI988/Rispens virus. PCR analysis and immunofluorenscence assays indicated that the recombinant virus was stable over 8 passages and that VP2 was expressed in CEF infected with the rCVI988-VP2. The data showed that the recombinant MDV expressing the VP2 gene of IBDV was successfully constructed and could prove to be a good strategy for IBD control.