Construction of plant expression vector containing human interferon gene and expression in Ginseng calli
- VernacularTitle:人干扰素基因植物表达载体的构建及其在人参愈伤组织细胞中的表达
- Author:
Qi REN
;
Jun SHENG
;
Zhixia REN
- Publication Type:Journal Article
- Keywords:
Human interferon-?2b;
Ginseng callus cells;
Transgene;
Agrobacterium tumefaciens
- From:
Chinese Journal of Immunology
2000;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express human interferon-?2b gene and to explore the feasibility of expressing human gene in plant cells.Methods:The hIFN-?2b coding sequence was amplified by PCR with specific primers and plasmid pBV889 was used as a template,subcloned into middle vector pMD18-T and binary vector pBI121 to obtain plant expression vector pBIFN. The pBIFN was transformed into Agrobacterium tumefaciens strain LBA4404. Then hIFN-?2b gene was introduced into Ginseng callus cells via Agrobacterium-mediated transformation. The positive cells were screened by G418. The transgenic Ginseng calli were confirmed by PCR,RT-PCR,Western blot and WISH/VSV system.Results:Stable integration of the hIFN-?2b gene in the Ginseng callus cells′ genome was confirmed by PCR analysis. RT-PCR analysis showed that there were transcription products. Western blot implied that the given protein was hIFN-?2b. WISH/VSV system assay showed that the expressed hIFN-?2b possessed relatively lower bioactivity.Conclusion:HIFN-?2b has been expressed in transgenic Ginseng calli, which facilitates further investigation of improving the curative effect of orally administered hIFN-?2b.
