人白细胞介素18和新城疫病毒HN基因嵌合表达载体的构建与表达  

Construction and expression of expresssion plasmid of hIL-18/NDV HN chimeric gene

作　　者：解丽华[1] 金宁一[2] 邵国喜[1] 米志强[2] 连海[2] 薛丽娟[2] 李萍[2] 

机构地区：[1]吉林大学第二医院肿瘤内科,吉林长春130041 [2]解放军军需大学病毒研究所解放军基因工程重点实验室,吉林长春130062

出　　处：《吉林大学学报（医学版）》2005年第3期352-354,共3页Journal of Jilin University：Medicine Edition

基　　金：国家 973项目资助课题 (G1990 1190 2 )

摘　　要：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 an eukaryotic expression plasmid containing the chimeric gene coding for the hemagglutinin-neuraminidase(HN) of newcastle disease virus(NDV) and human IL-18. Methods The enzymolysis and ligation were used to joint HN cDNA to IL-18 cDNA of pVAX1 IL-18 plasmid and at the same time the terminator of IL-18 cDNA was replaced, an eukaryotic expression vector of chimeric gene for HN and IL-18 was constructed. The recombinant plasmid was transfected into HeLa cells with liposome and the expression products were examined by hemagglutination test and Western blotting. Results The expression plasmid of pVAX1 IL-18 HN was contructed successfully confirmed by enzyme digestion identification. The expressions of IL-18 and HN chimeric genes were confirmed, and the expressed HN protein had higher hemagglutinative titer.Conclusion The constructed eukaryotic expression plasmid can express in vitro,and the expression products of the chimeric gene have good reactivity and specificity.

关 键 词：新城疫病毒 重组 遗传 白细胞介素18 嵌合表达质粒 基因表达 

分 类 号：Q78[生物学—分子生物学]