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glycoprotein e gene
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  glycoprotein e gene
Cloning and Regulation of the Promoter of Pseudorabies Virus (TNL Strain) Glycoprotein E Gene
      


A recombinant plasmid pSDM1. 78 + containing the complete glycoprotein E gene of pseudorabies virus (PRV) Ea strain was constructed by subcloning from plasmid pSKB4.5, which was composed of the gI, gE,11K genes and partial sequence of gD,28K genes. The results of sequence analysis showed that there were multiple mutants compared with PRV Rice strain. The gE gene was further cloned into the KpnI and BamHI sites of eukaryotic expression vector pcDNA3.1 + ,resuting in a eukaryotic expression...

A recombinant plasmid pSDM1. 78 + containing the complete glycoprotein E gene of pseudorabies virus (PRV) Ea strain was constructed by subcloning from plasmid pSKB4.5, which was composed of the gI, gE,11K genes and partial sequence of gD,28K genes. The results of sequence analysis showed that there were multiple mutants compared with PRV Rice strain. The gE gene was further cloned into the KpnI and BamHI sites of eukaryotic expression vector pcDNA3.1 + ,resuting in a eukaryotic expression plasmid pcDNA-gE. This expression plasmid was transfected into the IBRS-2 cell and the cell lines expressed stablely glycoprotein E was established. Expression protein distributed on the membrance and had biology activity by Indirect Immunoflures-ence Assay.

对含伪狂犬病病毒(Pseudorabies Virus,PrV)Ea株gD基因部分编码序列,gI、gE和11k基因全序列、28k基因部分序列的质粒pSKB4.5进行亚克隆,构建了只含完整gE基因(长1.78kb)的重组质粒pSDM1.78+,并采用双脱氧末端终止法对全序列进行了分析,发现同国外标准毒株Rice株相比较,在核苷酸和氨基酸水平均存在一定程度的差异。进一步将gE基因克隆到高效真核表达载体pcDNA3.1+的KpnI和BamHI位点之间,构建了gE基因的真核表达质粒pcDNA-gE。体外转染 IBRS-2细胞,经间接免疫荧光法检测证实了gE基因在IBRS-2细胞中得到了表达,表达的蛋白具有生物学活性。

Objective To construct the eukaryotic expression vector which express the protein of VZV glycoprotein E in COS7 cells.Methods VZV glycoprotein E genes were amplified with PCR and cloned into the eukaryotic expression vector pcDNA3.1.The constructed vector was transfected into COS7 cells with lipofection.The expressed VZV glycoprotein E was detected by means of immunohistochemistry.Results The amplified gene,which was about 1.9kb, included entire gene of VZV glycoprotein...

Objective To construct the eukaryotic expression vector which express the protein of VZV glycoprotein E in COS7 cells.Methods VZV glycoprotein E genes were amplified with PCR and cloned into the eukaryotic expression vector pcDNA3.1.The constructed vector was transfected into COS7 cells with lipofection.The expressed VZV glycoprotein E was detected by means of immunohistochemistry.Results The amplified gene,which was about 1.9kb, included entire gene of VZV glycoprotein E.The cells transfected with the constructed vector expressed VZV glycoprotein E successfully.Conclusion The constructed vector containing gene of VZV glycoprotein E can express glycoprotein E in COS7 cell.The success of constructing this vector laid the foundation for studying immunogenicity of glycoprotein E and VZV DNA vaccine.

目的 构建水痘 带状疱疹病毒 (VZV )糖蛋白E的真核表达载体并使其在COS7细胞中表达。方法 用PCR方法扩增VZV糖蛋白E基因 ,并将其克隆到真核表达载体 pcDNA 3 .1。用脂质体转染试剂将重组载体转入COS7细胞 ,通过免疫组化法检测转染细胞瞬时表达的重组蛋白。结果 扩增的目的基因包括了全段糖蛋白E基因 ,长度约1.9kb。重组表达载体在COS7细胞中表达出了糖蛋白E。结论 VZV糖蛋白E真核表达载体的成功构建及在COS7细胞中重组蛋白的表达 ,为研究此蛋白的免疫原性和VZV核酸疫苗打下了基础。

Objective To construct the eukaryotic expression vector of varicella-zoster virus(VZV)glycoprotein E.Methods VZV glycoprotein E genes were amplified by PCR and cloned into the eukaryotic expression vector pcDNA3.1.The constructed vector was identified by double enzyme-digestive method and sequencing.Results The amplified gene,which was about 1.9 kb,included entire gene of VZV glycoprotein E.The recombinant vector was identified as pcDNA-VgE.Conclusion The constructed...

Objective To construct the eukaryotic expression vector of varicella-zoster virus(VZV)glycoprotein E.Methods VZV glycoprotein E genes were amplified by PCR and cloned into the eukaryotic expression vector pcDNA3.1.The constructed vector was identified by double enzyme-digestive method and sequencing.Results The amplified gene,which was about 1.9 kb,included entire gene of VZV glycoprotein E.The recombinant vector was identified as pcDNA-VgE.Conclusion The constructed vector containing gene of VZV glycoprotein E can express glycoprotein E,which lays the foundation for studying immunogenicity of glycoprotein E.

目的构建水痘-带状疱疹病毒(VZV)糖蛋白E的真核表达载体。方法用PCR方法扩增VZV糖蛋白E基因,并将其克隆到真核表达载体pcDNA3.1并用双酶切和测序方法鉴定。结果扩增的目的基因包括了全长的糖蛋白E基因,长度约1.9 kb。并且成功构建了糖蛋白E基因重组表达载体。结论构建的重组表达载体基因序列正确,为进一步研究此蛋白的免疫原性打下了基础。

 
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