摘要
背景:葡聚糖结合蛋白B(gbpB)具有良好的免疫原性,但能否作为免疫防龋的候选基因疫苗有待研究。目的:观察gbpB真核表达质粒在哺乳动物细胞COS-7的表达情况。设计:设立对照的实验研究。地点和对象:实验在解放军第四军医大学口腔医学院牙体病科完成,对象为Pbs-gbpB质粒(本实验室构建)。干预:通过基因重组技术构建真核表达质粒pcDNA3.1(+)-gbpB,脂质体转染法将其转染至COS-7细胞。免疫组化SABC法检测,DAB显色。主要观察指标:转染后细胞胞浆、胞核着色情况。结果:pcDNA3.1(+)-gbpB质粒转染的细胞胞浆中呈棕黄色染色,胞核中无着色,pcDAN3.1(+)空载体转染的细胞胞浆及胞核无着色,空白对照组也无着色。结论:质粒pcDNA3.1(+)-gbpB转染COS-7后能够在细胞内翻译、表达,表达的蛋白质位于胞浆中,可与抗GbpB抗体特异性结合,具有抗原性,可作为基因疫苗。
BACKGROUND:Glucan binding protein B(gbpB) has good immunogenicity;however,it is still waiting for further researches to determine whether it can be selected as a candidate gene vaccine in immune prevention of caries. OBJECTIVE:To observe the expression of gbpB eukaryon expressed plasmid in the COS 7 cells of mammals. DESIGN:A control experimental study was conducted. SETTING and PARTICIPANTS:This study was completed in the Department of Operative Dentistry,College of Stomatology,Fourth Military Medical University.The subjects were Pbs gbpB plasmid established by our laboratory. INTERVENTIONS:Eukaryon expressed plasmid pcDNA3.1(+) gbpB was established by gene recombinant technique,which was transfected into COS 7 cells by liposome transfection method.The results were assayed by immunohistochemistry SABC method and colorated by DAB staining. MAIN OUTCOME MEASURES:The coloration of the cytoplasm and nucleus after transfection. RESULTS:The cytoplasm of the cells trasfected by pcDNA3.1(+) gbpB plasmid was in nankeen color and there was no staining in nucleus.There was no coloration in the cytoplasm and nucleus of the cells transfected by pcDNA3.1(+) empty carrier as well as that of control group. CONCLUSION:pcDNA3.1(+) gbpB can be translated and expressed in COS 7 cell after transfection.The expressive protein is in cytoplasm,which can connect with anti gbpB specific antibody with antigenicity;therefore,it can be used as a gene vaccine.
出处
《中国临床康复》
CSCD
2004年第14期2750-2751,F012,共3页
Chinese Journal of Clinical Rehabilitation
基金
全军"十五"重点课题(01Z089)~~
