CAJ | 학술논문

目的构建狂犬病病毒糖蛋白基因DNA真核表达质粒,并检测其免疫原性.方法用RT-PCR法扩增和分离CTN株狂犬病病毒糖蛋白基因,测序后克隆至pcDNA5.0载体,构建重组质粒pcDNA5.0-G,提取质粒,转化293T细胞,检测糖蛋白瞬时表达,并以该重组质粒肌肉注射免疫BALB/c小鼠,狂犬病病毒CVS株攻击,观察小鼠存活情况.结果酶切、测序结果显示重组质粒pcDNA5.0-G构建成功,瞬时表达结果显示糖蛋白获得大量表达.经肌肉注射质粒常规免疫小鼠,病毒攻击后小鼠保护率为73.3％,对照组为6.7％.结论所构建狂犬病病毒糖蛋白真核表达质粒pcDNA5.0-G经肌肉注射免疫后可有效保护小鼠免受狂犬病病毒攻击,具有良好的免疫原性,这为后期核酸疫苗的研发奠定基础.
목적 구건광견병병독당단백기인DNA진핵표체질립,병검측기면역원성.방법 용RT-PCR법확증화분리CTN주광견병병독당단백기인,측서후극륭지pcDNA5.0재체,구건중조질립pcDNA5.0-G,제취질립,전화293T세포,검측당단백순시표체,병이해중조질립기육주사면역BALB/c소서,광견병병독CVS주공격,관찰소서존활정황.결과 매절、측서결과현시중조질립pcDNA5.0-G구건성공,순시표체결과현시당단백획득대량표체.경기육주사질립상규면역소서,병독공격후소서보호솔위73.3％,대조조위6.7％.결론 소구건광견병병독당단백진핵표체질립pcDNA5.0-G경기육주사면역후가유효보호소서면수광견병병독공격,구유량호적면역원성,저위후기핵산역묘적연발전정기출.
Objective To construct the eukaryotic expression plasmid of the rabies virus glycoprotein gene DNA,and detect the immunogenicity.Methods Using RT-PCR amplified the glycoprotein gene of rabies virus CTN strain,sequenced and cloned into pcDNA5.0 (+) vector to construct the recombinant plasmid pcDNA5.0-G plasmid.Detect glycoprotein transient expression with transfecting the plasmid into 293T cells.Intramuscular immunization of BALB/c mice by the recombinant plasmid on day 0 and 7,then challenge by rabies virus CVS strain observed the mice survived.Results The results of the transient expression of glycoprotein abundantly expressed.The survival ratio of mice with CVS challenge after routine intramuscular injection of pcDNA5.0-G plasmid is 73.3％,and 6.7％ for the control group.Conclusion Rabies virus glycoprotein eukaryotic expression plasmid pcDNA5.0-G was successfully constructed,and has been good immunogenicity.It's to be the foundation for candidate DNA vaccine research and development.