CAJ | 학술논문

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(+)-gnpB,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.
배경:포취당결합단백B(gbpB)구유량호적면역원성,단능부작위면역방우적후선기인역묘유대연구.목적:관찰gbpB진핵표체질립재포유동물세포COS-7적표체정황.설계:설립대조적실험연구.지점화대상:실험재해방군제사군의대학구강의학원아체병과완성,대상위Pbs-gbpB질립(본실험실구건).간예:통과기인중조기술구건진핵표체질립pcDNA3.1(+)-gnpB,지질체전염법장기전염지COS-7세포.면역조화SABC법검측,DAB현색.주요관찰지표:전염후세포포장、포핵착색정황.결과:pcDNA3.1(+)-gbpB질립전염적세포포장중정종황색염색,포핵중무착색,pcDAN3.1(+)공재체전염적세포포장급포핵무착색,공백대조조야무착색.결론:질립pcDNA3.1(+)-gbpB전염COS-7후능구재세포내번역、표체,표체적단백질위우포장중,가여항GbpB항체특이성결합,구유항원성,가작위기인역묘.
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 tbe 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 tbe 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.