Preliminary Study on Expression Infectious Bronchitis Virus Level of the S1 Gene of Avian in a Baeulovirus System

SONG Yan-Hua; LIU Fu-An

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June 1999

SONG Yan-Hua and LIU Fu-An. Preliminary Study on Expression Infectious Bronchitis Virus Level of the S1 Gene of Avian in a Baeulovirus System[J]. Virologica Sinica, 1999, 14(3): 255-258.

Citation: SONG Yan-Hua, LIU Fu-An. Preliminary Study on Expression Infectious Bronchitis Virus Level of the S1 Gene of Avian in a Baeulovirus System .VIROLOGICA SINICA, 1999, 14(3) : 255-258.

鸡传染性支气管炎病毒s1基因在昆虫细胞中表达水平初探

宋延华 ,
刘福安

1.
华南农业大学动物医学系，广州510642

摘要

将含有鸡传染性支气管炎病毒Ｓ１基因ｃＤＮＡ的重组转移质粒ｐＳＸＩＶＶＩ＋Ｘ３Ｓ１．Ｈｏｌｔｅ和ｐＳＸＩＶＶＩ＋Ｘ３／４Ｓ１．Ｈｏｌｔｅ分别与粉纹夜蛾核型多角体病毒ＴｎＮＰＶＳＶＩ－ＧＤＮＡ（ＯＣＣ－，ｇａｌ＋）共转染草地夜蛾（Ｓｆ９）细胞，经空斑纯化得到重组病毒ＴｎＮＰＶ（Ｘ３）Ｓ１．ＨｏｌｔｅＯＣＣ＋和ＴｎＮＰＶ（Ｘ３／４）Ｓ１．ＨｏｌｔｅＯＣＣ＋。将重组毒株分别感染Ｔｎ５Ｂ１细胞，并进行ＳＤＳＰＡＧＥ与Ｗｅｓｔｅｒｎｂｌｏｔ检测。结果表明，ＴｎＮＰＶ（Ｘ３／４）Ｓ１．ＨｏｌｔｅＯＣＣ＋在感染的细胞中高效表达了Ｓ１蛋白，ＳＤＳＰＡＧＥ凝胶薄层色谱分析结果显示，感染病毒后７２ｈＳ１蛋白的表达量占细胞内总蛋白量的３５．８％，而ＴｎＮＰＶ（Ｘ３）Ｓ１．ＨｏｌｔｅＯＣＣ＋感染的细胞内检测不出Ｓ１蛋白。经分析认为这一差异主要来自Ｓ１基因翻译起始位点及其附近的周围环境。
- 传染性支气管炎病毒
- , S1基因
- , 杆状病毒表达载体
- , 表达

Preliminary Study on Expression Infectious Bronchitis Virus Level of the S1 Gene of Avian in a Baeulovirus System

SONG Yan-Hua ,
LIU Fu-An

1.
Oepartmen~of Veterinary Medicine，South China Agricutturat University，Guangzhou 510642

Abstract

The recombinant transfer plasmids pSXIVVI\++X3 S1.Holte and pSXIVVI\++X3/4 S1.Holte containing S1 gene cDNA of avian infectious bronchitis virus and the parent virus (TnNPV SVI -G) DNA were used to co transfect \%Spodotera frugiperda\% (Sf) cells. The recombinant viruses TnNPV (X3)S1.Holte OCC + and TnNPV (X3/4)S1.Holte OCC +, which included the S1 gene and could form polyhedra, were plaque purified. The expression products of S1 gene were detected by SDS PAGE and Western blotting in Tn 5B1 cells infected with the recombinant viruses. The S1 protein with a molecular weight of 100 kD was expressed in the cells infected with TnNPV (X3/4)S1.Holte OCC + at high efficiency, the expression level in Tn 5B1 cells 72 h postinfection being 35.8% by SDS PAGE gel thin layer chromatography. No expression product was detected by SDS PAGE and Western blotting in Tn 5B1 cells infected with TnNPV (X3)S1.Holte OCC +. The expression level depends mainly upon the translational start site of the S1 gene
- Avian infectious bronchitis virus （IBV）
- , S1gene
- , Baculovirus expression vector
- , Expression

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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

Preliminary Study on Expression Infectious Bronchitis Virus Level of the S1 Gene of Avian in a Baeulovirus System

SONG Yan-Hua ,
LIU Fu-An

1. Oepartmen~of Veterinary Medicine，South China Agricutturat University，Guangzhou 510642

Abstract: The recombinant transfer plasmids pSXIVVI\++X3 S1.Holte and pSXIVVI\++X3/4 S1.Holte containing S1 gene cDNA of avian infectious bronchitis virus and the parent virus (TnNPV SVI -G) DNA were used to co transfect \%Spodotera frugiperda\% (Sf) cells. The recombinant viruses TnNPV (X3)S1.Holte OCC + and TnNPV (X3/4)S1.Holte OCC +, which included the S1 gene and could form polyhedra, were plaque purified. The expression products of S1 gene were detected by SDS PAGE and Western blotting in Tn 5B1 cells infected with the recombinant viruses. The S1 protein with a molecular weight of 100 kD was expressed in the cells infected with TnNPV (X3/4)S1.Holte OCC + at high efficiency, the expression level in Tn 5B1 cells 72 h postinfection being 35.8% by SDS PAGE gel thin layer chromatography. No expression product was detected by SDS PAGE and Western blotting in Tn 5B1 cells infected with TnNPV (X3)S1.Holte OCC +. The expression level depends mainly upon the translational start site of the S1 gene