单纯疱疹病毒糖蛋白D与IL-2基因共表达质粒的构建及其免疫学鉴定  

Co-expression of HSV-1 gD gene and interleukin-2 gene in dendritic cell

作　　者：李伟荣[1] 王得新[1] 王健伟[2] 牛勃 胥显民 

机构地区：[1]首都医科大学附属北京友谊医院神经内科,100050 [2]中国疾病预防控制中心病毒病预防控制所 [3]北京北医联合生物医学中心

出　　处：《中华实验和临床病毒学杂志》2007年第4期328-330,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的构建含单纯疱疹病毒Ⅰ型糖蛋白D(gD)全长基因和人白细胞介素2(IL-2)共表达的重组真核表达质粒IRES-gD-IL-2,并诱导其在抗原提呈细胞---树突状细胞(DCs)的表达。方法应用PCR方法分别扩增出HSV-1gD和IL-2基因,经PCR、双酶切及测序证实正确后分别插入真核表达载体IRES的上、下游。通过脂质体介导转染DCs,并进行免疫学鉴定。结果重组表达质粒IRES-gD-IL-2经PCR和酶切均出现相应长度的片段。经Western Blot证实,HSV-1gD抗体可特异识别DCs内表达的gD蛋白。结论构建了gD与IL-2共表达的重组真核表达质粒IRES-gD-IL-2,免疫印迹鉴定证实该表达产物具有免疫学活性。Objective To construct eukaryotic expression plasmid IRES-gD-IL-2 which contains both HSV-1 glycoprotein D （gD） gene and IL-2 gene and to induce it to express in antigen presenting cell （APC）-- dendritic cells （ DCs ）. Methods The whole sequence of gD and IL-2 were amplified by PCR assay. After confirmation by PCR, double-enzyme digestion and sequencing, these genes were directly cloned into eukaryotic expression vector IRES, then were transfected into DCs. Western blotting was employed to identify the transcription and expression of gD gene. Results The results of PCR and enzyme digestion showed that the recombinant expression plasmid contained correct fragments, and the transcription and expression of gD were cotffirmed by Western blotting. Conclusion The recombinant expression vector IRES-gD-IL-2 was constructed, the results of the Western blotting showed that the recombinant protein could be identified by gD- specific antibody, therefore the protein has immunologic competence.

关 键 词：疱疹病毒1型 人 糖蛋白类 白细胞介素2 

分 类 号：R346[医药卫生—基础医学] R392