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

Study on the cloning of hantan virus glycoprotein G2 gene and its immunogenicity

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。

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 this plasmid and the anti-HTNV 76～118 strain cross-antibodies were detected by IFA. Results Cross-antibodies against hantavirus could be tested in the sera of the immunized mice, and the titers were 1:10.99, which showed significantly difference with the control. Conclusions The results suggest that the fusion gene pcDNA 3.1/HisB-G2 is successively constructed and it can directly stimulate BALB/C mice to produce specific humoral immunity, but the titers are not very high. This study provides an experimental basis for the future research of efficient DNA vaccine for HFRS.