The recombinant plasmid pBIBSa containing the HBsAg DNA fragment was transferred into Agrobacterium tumefaciens strain LBA4404 directly. Ginseng cells were transfected with A. tumefaciens carrying pBIBSa and the ginse...The recombinant plasmid pBIBSa containing the HBsAg DNA fragment was transferred into Agrobacterium tumefaciens strain LBA4404 directly. Ginseng cells were transfected with A. tumefaciens carrying pBIBSa and the ginseng cell lines carrying HBsAg-S gene were obtained. The presence of target gene in the transfect cells was confirmed by PCR and RT-PCR. A clear band at the site of 700 bp was observed by agarose electrophoresis analysis of the samples containing the target gene. HBsAg expressed by the transgenic ginseng cells was detected by Western blot. Maximum expression levels of 184 ng HBsAg/g FW and 0. 009% of the total soluble proteins were observed by ELISA. HBsAg in ginseng cells was located both on the cell membrane and in the nuclei.展开更多
基金Supported by the Science and Technology Development Planning Foundation of Jilin Province of China(No.20030405)
文摘The recombinant plasmid pBIBSa containing the HBsAg DNA fragment was transferred into Agrobacterium tumefaciens strain LBA4404 directly. Ginseng cells were transfected with A. tumefaciens carrying pBIBSa and the ginseng cell lines carrying HBsAg-S gene were obtained. The presence of target gene in the transfect cells was confirmed by PCR and RT-PCR. A clear band at the site of 700 bp was observed by agarose electrophoresis analysis of the samples containing the target gene. HBsAg expressed by the transgenic ginseng cells was detected by Western blot. Maximum expression levels of 184 ng HBsAg/g FW and 0. 009% of the total soluble proteins were observed by ELISA. HBsAg in ginseng cells was located both on the cell membrane and in the nuclei.
