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目的:构建人白细胞介素18(IL - 18)与新城疫病毒(NDV) HN基因嵌合表达质粒。方法:选用适当的限制性核酸内切酶将NDV的HN基因连接至真核表达质粒p VAX1IL - 18中IL - 18基因的尾部,同时替换掉IL - 18基因的终止子,构建表达IL - 18HN嵌合基因的p VAX1IL - 18HN质粒,经酶切鉴定正确后,通过脂质体介导转染He L a细胞,应用Western blotting及血凝试验检测其表达情况。结果:酶切鉴定证实正确地构建了p VAX1IL - 18HN嵌合表达质粒,Western blotting和血凝试验检测结果表明,嵌合后的IL - 18和HN基因均能够表达,且表达的HN蛋白具有较高的血凝活性。结论:IL - 18和HN基因嵌合后不影响二者的表达,而且表达的HN蛋白具有血凝活性。
Objective: To construct the chimeric expression plasmid of human interleukin 18 (IL-18) and Newcastle disease virus (NDV) HN gene. METHODS: The appropriate restriction endonuclease was used to connect the NDV HN gene to the tail of the IL-18 gene in the eukaryotic expression plasmid pVAX1IL-18. At the same time, the terminator of the IL-18 gene was replaced to construct the IL-18HN. The p VAX1IL-18HN plasmid of chimeric gene was identified by restriction enzyme digestion and then transfected into He L a cells by liposome. Western blotting and hemagglutination assay were used to detect its expression. RESULTS: Enzyme digestion analysis confirmed that the pVAX1IL-18HN chimeric plasmid was constructed correctly. Western blotting and hemagglutination tests showed that the chimeric IL-18 and HN genes could all be expressed and the expressed HN protein had better results. High blood clotting activity. CONCLUSION: IL-18 and HN gene chimerism do not affect the expression of both genes, and the expressed HN protein has hemagglutination activity.