Cloning of Gene Encoding G Protein from Respiratory Syncytial Virus and Co-expression of G Protein Fragments with Diverse Carrier Proteins

FAN Chang-fa; GONG Wei; MEI Xing-guo; LIU Yan

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December 2004

FAN Chang-fa, GONG Wei, MEI Xing-guo and LIU Yan. Cloning of Gene Encoding G Protein from Respiratory Syncytial Virus and Co-expression of G Protein Fragments with Diverse Carrier Proteins[J]. Virologica Sinica, 2004, 19(6): 553-558.

Citation: FAN Chang-fa, GONG Wei, MEI Xing-guo, LIU Yan. Cloning of Gene Encoding G Protein from Respiratory Syncytial Virus and Co-expression of G Protein Fragments with Diverse Carrier Proteins .VIROLOGICA SINICA, 2004, 19(6) : 553-558.

呼吸道合胞病毒G蛋白基因片段的克隆及高效表达

范昌发 ,
梅兴国 ,

1.
军事医学科学院毒物药物研究所，北京

摘要

呼吸道合胞病毒 (RSV)感染遍布全球，并可导致严重的疾病，但目前尚无成功的疫苗问世。为寻求可能用于RSV疫苗研制的重组蛋白抗原，我们在克隆RSV-A全长G蛋白基因的基础上，构建了多种共表达载体蛋白和G蛋白片段的表达载体，并从中筛选出能以可溶形式高效表达抗原蛋白的原核表达体系。通过亲和层析纯化了重组蛋白抗原DsbA-G101，将其免疫Balb/c小鼠后获得了相应的抗血清。经ELISA检测表明DsbA-G101具有良好的免疫原性。基于本研究所构建的系列表达载体，可以比较不同的G蛋白片段免疫原性的强弱及载体蛋白的优劣，从中发现最佳的RSV抗原蛋白。
- RSV
- , G蛋白
- , 载体蛋白
- , 表达
- , 疫苗

Cloning of Gene Encoding G Protein from Respiratory Syncytial Virus and Co-expression of G Protein Fragments with Diverse Carrier Proteins

1.
Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, 27 Taiping Road, Beijing 100850, China

Corresponding author: MEI Xing-guo,

Abstract

Although severe diseases may be caused by respiratory syncytial virus (RSV) infection in the world, no efficacious vaccine against this virus is licensed. In an effort to seek recombinant protein antigens that may be used in the future vaccine development, we constructed a series of expression vectors which can co-express carrier proteins and G protein fragments based on cloning the full cDNA sequence of G protein from RSV. An expression system that can efficiently express recombinant protein antigens in soluble form was selected for subsequent researches. Balb/c mice were immunized with one of the purified recombinant protein antigens, DsbA-G101, for production of anti- serum. Based on ELISA assays, good immunogenicity of DsbA-G101 was revealed. The constructed expression vectors can be used in immunogenicity analysis of different G protein fragments and in selection of ideal carrier proteins, two aspects which are important for the future development of vaccine against this virus.
- Respiratory syncytial virus
- , G protein
- , Carrier protein
- , Expression
- , 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

Cloning of Gene Encoding G Protein from Respiratory Syncytial Virus and Co-expression of G Protein Fragments with Diverse Carrier Proteins

Corresponding author: MEI Xing-guo,

1. Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, 27 Taiping Road, Beijing 100850, China

Abstract: Although severe diseases may be caused by respiratory syncytial virus (RSV) infection in the world, no efficacious vaccine against this virus is licensed. In an effort to seek recombinant protein antigens that may be used in the future vaccine development, we constructed a series of expression vectors which can co-express carrier proteins and G protein fragments based on cloning the full cDNA sequence of G protein from RSV. An expression system that can efficiently express recombinant protein antigens in soluble form was selected for subsequent researches. Balb/c mice were immunized with one of the purified recombinant protein antigens, DsbA-G101, for production of anti- serum. Based on ELISA assays, good immunogenicity of DsbA-G101 was revealed. The constructed expression vectors can be used in immunogenicity analysis of different G protein fragments and in selection of ideal carrier proteins, two aspects which are important for the future development of vaccine against this virus.