汉坦病毒囊膜蛋白基因的克隆及其免疫原性的初步研究.pdfVIP

5·48 · Chin J Dis Cont rol Prev 　2004 　Dec ; 8 (6) 汉坦病毒 G2 囊膜蛋白基因的克隆及其免疫原性的初步研究 黄汉菊 , 吴志洪 , 王小红 【摘要】　目的 　构建汉坦病毒 G2 囊膜蛋白基因真核表达载体 ,进一步研究其免疫效果。方法 　 采用 PCR 方法扩增出 G2 基因 ,亚克隆于 pcDNA3. 1/ HisB 载体 ,重组阳性克隆进行酶切和测序鉴 ( ) 定。将重组质粒免疫 BALB/ C 小鼠 ,用间接免疫荧光法 IFA 检测免疫小鼠血清中抗汉坦病毒 76～118株的交叉抗体。结果 　5 只免疫小鼠血清中均检测到特异性的抗汉坦病毒 76～118 株的 交叉抗体 ,与对照组相比有显著性差异 ,其滴度为 1 ∶10. 99 。结论 　G2 囊膜蛋白基因构建成功并 能刺激机体产生特异性抗体 ,但其效价不高 ,该研究结果为研制有效的 HFRS 核酸疫苗奠定一定 的实验基础。 【关键词】　汉坦病毒 ; 基因 【中图分类号】R373. 9 　【文献标识码】A 　【文章编号(2004) Study on the cloning of hantan virus glycoprotein G2 gene and its immunogenicity 　HUAN G Hanju , WU Zhihong , WAN G Xiaohong. 　Depart ment of Pathogenetic Biology , School of B asic Medical Sciences , Tongji Medical College , Huaz hong U niversity of Science and Technology , W uhan 　430030 , China 【Abstract 】　Objective 　To construct the eukaryotic expression vector of G2 gene of hantavirus envelope glycoprotein and to study its immunization effect. Methods 　The G2 gene was amplified by PCR and cloned into the eukaryotic expression vector pcDNA3. 1/ HisB , verified the recombinant with restriction endonuclease and sequenced. The BALB/ C mice were immunized with thisplasmid and the antiHTNV 76～ 118 strain crossantibodies were detected by IFA. Results 　Crossantibodies against hantavirus could be tested in the sera of the immunized mice , and the titers were 1 :10. 99 , which showed signifi cantly difference with the control. Conclusions 　The results suggest that