YANG Yi—liang, LIANG Guo—dong, FU Shi—hong, SU Nai—lun, DENG Juan and HOU Yun—de. Construction of Replicon Expression Vector Derived from XJ-160 Virus[J]. Virologica Sinica, 2003, 18(3): 221-226.
Citation: YANG Yi—liang, LIANG Guo—dong, FU Shi—hong, SU Nai—lun, DENG Juan, HOU Yun—de. Construction of Replicon Expression Vector Derived from XJ-160 Virus .VIROLOGICA SINICA, 2003, 18(3) : 221-226.

XJ一160病毒复制子型表达载体的构建

  • xJ-160病毒是我国首次分离的辛德毕斯病毒,全基因组测序已经完成。本文利用该病毒全基因序列首先构 建了全基因组cDNA克隆质粒,在此基础上,利用基因重组技术将病毒结构基因序列替换为含有多个单酶切位点 的序列,得到复制子表达载体质粒pRepxj160。为验证载体的功能,将报告基因绿色荧光蛋白(EGFP)和B一半 乳糖苷酶基因(1acZ)分别插入到载体的多克隆位点,得到两个表达质粒;经体外转录获得的转录体RNA 转染 BHK-21细胞后14h,可检测到报告基因的表达。结果表明我们构建的XJ.160病毒复制子型表达载体具有自主复 制功能,可以表达异源基因。本研究为进一步开发具有我国自主知识产权的甲病毒载体奠定了基础。

Construction of Replicon Expression Vector Derived from XJ-160 Virus

  • Abstract:XJ一1 60 is the first Chinese isolate of a Sindbis—like virus,an d its genome was completely sequenced in our previous work.In this investigation,a full—length genomic cDNA clone Was firstly constructed from RT—PCR products of the viral RNA.Then.the plasmid of replicon expression vector (pRepxj 1 60)was derived from this cDNA clone replacin~viral structural sequence with multiclonal seq uence by DNA recombination technique.To verify function of this vector,reporter genes.EGFP an d cZ.were cloned into this plasmid.respectively.And RNA tran scripted from expression plasmid Was inducted into BHK一2 l cell line. resulting in expression of green fluorescent protein an d 13 .galactosidase 14 hours later.Th e results indicated mat the replicon vector derived from )(J—l60 virus was self-replicating and that the following gene expression was efficient.Our study settled the basis for developing all:Ihavirus vector system with Chinese intellectual property.

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    Construction of Replicon Expression Vector Derived from XJ-160 Virus

    • 1. National Institute For Viral Disease Control and Prevention,Chinese Cenwr For Disease Control And Prevention, Beifing 100052,China

    Abstract: Abstract:XJ一1 60 is the first Chinese isolate of a Sindbis—like virus,an d its genome was completely sequenced in our previous work.In this investigation,a full—length genomic cDNA clone Was firstly constructed from RT—PCR products of the viral RNA.Then.the plasmid of replicon expression vector (pRepxj 1 60)was derived from this cDNA clone replacin~viral structural sequence with multiclonal seq uence by DNA recombination technique.To verify function of this vector,reporter genes.EGFP an d cZ.were cloned into this plasmid.respectively.And RNA tran scripted from expression plasmid Was inducted into BHK一2 l cell line. resulting in expression of green fluorescent protein an d 13 .galactosidase 14 hours later.Th e results indicated mat the replicon vector derived from )(J—l60 virus was self-replicating and that the following gene expression was efficient.Our study settled the basis for developing all:Ihavirus vector system with Chinese intellectual property.

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