猪囊尾蚴抗原与猪白细胞介素-4基因融合DNA疫苗载体的构建

CONSTRUCTION OF DNA VACCINE INCLUDING A CHIMERIC GENE ENCODING CYSTICERCUS CELLULOSAE ANTIGEN AND PORCINE INTERLEUKIN 4

目的： 为增强猪囊尾蚴抗原ｃ Ｃ１ 的免疫保护作用而构建一表达猪白细胞介素４（ Ｉ Ｌ４）与 ｃ Ｃ１ 抗原融合蛋白的 Ｄ Ｎ Ａ 疫苗载体。方法： 通过聚合酶链反应（ Ｐ Ｃ Ｒ），分别扩增猪 Ｉ Ｌ４ｃ Ｄ Ｎ Ａ 和ｃ Ｃ１ｃ Ｄ Ｎ Ａ 片段并进行融合，获得的嵌合基因 Ｉ Ｌ４ｃ Ｃ１ 中含有１０ 个氨基酸的中间接头序列，同时对其５’ Ａ Ｕ Ｇ 侧翼序列进行翻译优化突变。结果：经酶切鉴定，证实有一１．５ ｋｂ 的片段插入载体，其 Ｄ Ｎ Ａ 序列分析结果与文献报道和实验设计完全一致。结论：表达猪 Ｉ Ｌ４ 与猪囊尾蚴抗原ｃ Ｃ１ 融合蛋白的 Ｄ Ｎ Ａ 疫苗载体构建成功。

AIM: To construct a fusion expression vector for DNA vaccine including porcine interleukin 4(IL 4)and antigen cC1 to enhance the protective immunity of Cysticercus cellulosae antigen cC1. METHODS: The cDNA fragments encoding porcine IL 4 and cC1 were amplified respectively by PCR and then fused.The obtained chimeric gene IL 4cC1 contained a synthetic linker of ten amino acids and the sequence surrounding its 5’ AUG initiatory codon was changed to optimized translational initiation. RESULTS: Identified by restriction enzyme analysis,an insert fragment of 1.5 kb was demonstrated. It had the same sequence as reported and designed by DNA sequencing analysis.CONCLUSION: A fusion expression plasmid containing porcine IL 4 and cC1 was constructed.