人参愈伤组织细胞系表达乙肝病毒表面抗原的研究

发布时间：2018-02-03 01:37

本文关键词： 乙肝表面抗原人参愈伤组织细胞细胞培养　出处：《吉林大学》2005年硕士论文　论文类型：学位论文

【摘要】：为了能够获得更加经济、高效、安全的乙肝疫苗,本实验室运用根瘤农杆菌转染的方法,将乙肝表面抗原(HBsAg)基因转入人参愈伤组织细胞,筛选出稳定生长的转化体细胞,命名为CS83 细胞系。本研究分别应用ELISA、Western blotting 和免疫组织化学的方法,对CS83 细胞表达的HBsAg 进行了鉴定和细胞内定位。小鼠的注射免疫和家兔的口服免疫实验都证明,该细胞系表达的HBsAg 在动物体内能有效地诱导免疫应答,产生特异性抗体。同时发现,人参愈伤组织细胞内可能存在着某些能起到免疫佐剂作用的物质。在优化CS83 细胞固体培养条件的同时,进行了细胞的液体化培养,建立起了分散程度较好的悬浮细胞系,为CS83 细胞的工业化生产奠定了基础。本研究为利用植物组织培养技术生产医用蛋白提供了技术平台,为其他药用植物的开发利用奠定了坚实的基础。
[Abstract]:In order to obtain more economical, efficient and safe hepatitis B vaccine, Agrobacterium tumefaciens was used to transfect hepatitis B surface antigen HBsAg gene into ginseng callus cells. A stable growth transforming somatic cell line was selected and named as CS83 cell line. In this study, ELISAL Western blotting and immunohistochemical methods were used respectively. The expression of HBsAg in CS83 cells was identified and localized in cells. Both the injection immunization of mice and the oral immunization of rabbits were proved. The HBsAg expressed by the cell line can effectively induce immune response and produce specific antibodies in animals. There may be some substances in callus of ginseng which can act as immune adjuvant. The liquid culture of CS83 cells was carried out at the same time as optimizing the conditions of solid culture. A suspension cell line with good dispersion was established, which laid a foundation for the industrial production of CS83 cells. This study provides a technical platform for the production of medical proteins by using plant tissue culture technology and lays a solid foundation for the development and utilization of other medicinal plants.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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