摘要
目的构建SARSS抗原的B细胞表位核酸疫苗并观察其免疫效果。方法将已经鉴定的SARSS抗原的B细胞表位S603-620的编码DNA序列插入真核表达载体PCI中,构建核酸疫苗(PCI-S603-620)。使用EcoR Ⅰ和Xba Ⅰ酶切,电泳及DNA测序鉴定。使用脂质体将PCI-S603-620转入293A细胞,使用ELISA检测其在真核细胞中的表达。使用PCI-S603-620免疫小鼠,ELISA检测其诱导的抗体情况。结果对PCI-S603-620进行酶切鉴定,电泳结果表明目的基因片段分子量为100bp左右。PCI-S603-620的DNA测序结果表明其序列与设计序列吻合。PCI-S603-620能在转染的293A细胞中表达。PCI-S603-620免疫小鼠可以诱导特异性抗体产生,效价为1:400。该抗体可以特异性识别SARSS抗原。结论成功构建SARSS抗原的B细胞表位核酸疫苗(PCI-S603-620)。
Objective To design B-Cell epitope DNA vaccine against S protein of severe aeute respiratory syndrome coronavirus （SARS-CoV） and evaluate its immune effect in BLAB/c mice. Methods SARS S antigen B-Cell epitope S603-620 coding DNA sequence was cloned into eukaryotic expression vector PCI to construct recombinant DNA Vaccine（PCI-S60-620）. It was verified by restrietion enzymes cutting, electrophoresis experiment and DNA sequencing.The recombinant plasmid was transfected into 293A cells with liposome. ELISA was used to detect its expression. BLAB/c mice were inoculated with PCI-S^6~. The antibody was determined by ELISA. Results Restriction analysis enzymes cutting and electrophoresis experiment proved that target gene segment was about 100 bp. DNA sequeneing showed that the sequenee is the same with the designed sequence. PCI-S603-620 could express in transfeeted 293A cells. The titer of antibody it indueed in BLAB/c mice was 1：400, which could specifically recognize SARS S antigen. Conclusion B-Cell epitope DNA vaccine against S protein of SARS was suceessfully eonstrueted.
出处
《热带医学杂志》
CAS
2012年第7期813-816,共4页
Journal of Tropical Medicine
基金
广州市珠江科技新星项目(2011022)
广州医学院博士启动基金(0706054)
