[Expression of hepatitis B surface antigen gene in ginseng cell].
The recombinant plasmid pBIBSa containing the HBsAg DNA fragment was transferred into Agrobacterium tumefaciens strain LBA4404 directly. Ginseng cells were co-cultivated with Agrobacterium tumefaciens carrying pBIBSa. The ginseng cell lines carrying HBsAg-S gene were obtained. Transgenic cells were checked for the presence of target gene using PCR and RT-PCR. Samples containing the target gene showed a clear band at the site of 700 bp by agarose electrophoresis analysis (Figs. 2, 3). Expression levels determined by ELISA showed maximum expression levels of 184 ng HBsAg/g FW and 0.009% of the total soluble protein. HBsAg in ginseng cells were located both on the membrane and in the nuclei (Fig. 4).
Liu
Dan
D
Changchun Institute of Biological Product, Changchun 130062, China.
Yu
Hai-Peng
HP
Sheng
Jun
J
Liu
Xiao-Yu
XY
Wang
Zhi-Wu
ZW
Li
Juan
J
Sheng
Wei-Jin
WJ
Gao
Zheng-Lun
ZL
Yao
Wei-Min
WM
Cai
Wei-Ming
WM
D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000378:metabolism; D005894:Panax / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
D006514:Hepatitis B Surface Antigens