[Establishment of stably expressed human RANTES gene in prunella vulgaris cell clone].
To express interesting human genes in herbal cells for boosting their specific pharmacological activities, RANTES gene cloned from human peripheral blood lymphocyte (PBL) mRNA was introduced into A. tumefaciens strain LBA4404 harboring pAL4404 plasmid via tumor-inducing (Ti) plasmid-derived intermediate expression vector pROKII. In vitro cultured P. vulgaris cells were transformed by leaf-disk cocultivation procedure. Integration of RANTES gene in the genome of transformed cells was confirmed by Southern blotting, and expression of RANTES gene in transformed cells was analyzed by RT-PCR amplification, Western blotting and enzyme-linked immunosorbent assay (ELISA). The peroxidase activity of PBL was utilized as a detection index of cellular chemotropism induction by recombinant RANTES. The results have shown the RANTES gene was integrated in transgenic P. vulgaris cells, and RANTES gene-stably expressed cell clones were available, which could pave the way to obtain transgenic P. vulgaris plants demonstrating specific pharmacological activities.
Zeng
Qing-Ping
QP
Biotechnology Laboratory, Tropical Medicine Institute, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, China. tmibio@gzhtem.edu.cn
Feng
Li-Ling
LL
Yang
Rui-Yi
RY
Chen
Zhu-Hua
ZH
D016960:Agrobacterium tumefaciens / Q000235:genetics; D015153:Blotting, Western; D018946:Chemokine CCL5 / Q000235:genetics / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D031347:Prunella / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018946:Chemokine CCL5