CAJ | 학술논문

目的利用293T细胞表达、纯化圣路易斯脑炎病毒样颗粒(Virus like particles,VLPs),为研制圣路易斯脑炎病毒免疫诊断试剂奠定基础.方法构建含圣路易斯脑炎病毒PrM和E基因的重组质粒pcDNA5/FRT-SJME,瞬时转染293T细胞,表达并纯化重组蛋白,通过透射电镜、间接免疫荧光试验、Western-Blot和ELISA对其进行鉴定.结果表达的重组蛋白形成50 nm的球型颗粒,能与抗圣路易斯脑炎病毒抗体产生特异性反应,与部分黄病毒属阳性血清发生交叉反应.结论已成功在哺乳动物细胞中表达并纯化了圣路易斯脑炎病毒样颗粒,其具有良好的抗原性和完整的颗粒形态,为进一步研制圣路易斯脑炎病毒感染的免疫诊断试剂奠定了基础.
목적 이용293T세포표체、순화골로역사뇌염병독양과립(Virus like particles,VLPs),위연제골로역사뇌염병독면역진단시제전정기출.방법 구건함골로역사뇌염병독PrM화E기인적중조질립pcDNA5/FRT-SJME,순시전염293T세포,표체병순화중조단백,통과투사전경、간접면역형광시험、Western-Blot화ELISA대기진행감정.결과 표체적중조단백형성50 nm적구형과립,능여항골로역사뇌염병독항체산생특이성반응,여부분황병독속양성혈청발생교차반응.결론 이성공재포유동물세포중표체병순화료골로역사뇌염병독양과립,기구유량호적항원성화완정적과립형태,위진일보연제골로역사뇌염병독감염적면역진단시제전정료기출.
Objective To express St.Louis encephalitis virus-like particles in mammalian cells,it will provide a prerequisite for further immunological diagnostic studies.Methods 293T-cell were transiently transfected with recombinant PreM-E plasmid.Expression and antigenicity of the purified protein were determined by transmission electron microscope (TEM),Western-Blot,immunofluorescence assay and ELISA.Results Recombinant subviral particles,about 50 nm in diameter,were observed by TEM in the supernatant of transfected cells.The results of Westeru-Blot,IFA and ELISA showed the recombinant proteins retained immunoreactivity similar to those of native virus proteins.Conclusion St.Louis encephalitis viruslike particles has good antigenicity and physical appearance,It will provide a prerequisite for further immune diagnostic reagent.