ABSTRACT
Phytoene synthase is a rate-limiting enzyme in the pathway of carotenoid biosynthesis.To aim at establishing a transformation of Ginseng callus cells, elevating the nutritive value through encouraging the composition of corresponding carotenoid,taking Ginseng callus cells as acceptor,psy gene were transformed into cells via Agrobacterium-mediated. The factors affecting genetict ransformation were also investigated seperately from concentrations of A.tumefaciens, age of Ginseng callus cells, infection time and co-culture time. PCR,PCR-Southern and RT-PCR analysis from the transferred gene plants proved that psy gene has been transferred into Ginseng callus cells and can be expressed. The content of ?-carotene was increased by an average of 26 times.A base of improving and raising the contents of carotenoid in the Ginseng callus cell was established.
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.