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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 construct the B cell epitope DNA vaccine of SARSS antigen and observe its immune effect. Methods The coding DNA sequence of the B cell epitopes S603-620 of the identified SARSS antigen was inserted into the eukaryotic expression vector PCI to construct a nucleic acid vaccine (PCI-S603-620). EcoR Ⅰ and Xba Ⅰ digestion, electrophoresis and DNA sequencing. PCI-S603-620 was transfected into 293A cells using liposomes and its expression in eukaryotic cells was assayed by ELISA. The mice were immunized with PCI-S603-620 and the induced antibodies were detected by ELISA. Results PCI-S603-620 digestion identification, electrophoresis results showed that the molecular weight of the target gene was about 100bp. The result of DNA sequencing of PCI-S603-620 shows that the sequence matches the designed sequence. PCI-S603-620 can be expressed in transfected 293A cells. PCI-S603-620 immunized mice can induce specific antibody production, titer of 1: 400. The antibody specifically recognizes the SARSS antigen. Conclusion The B cell epitope DNA vaccine of SARSS antigen was successfully constructed (PCI-S603-620).