Articles
STUDY ON TRANSFORMATION OF CYSTEINE PROTEINASE INHIBITOR GENE INTO CABBAGE (BRASSICA OLERACEA VAR. CAPITATA L.)
Article number
706_27
Pages
231 – 238
Language
English
Abstract
The plant expression vector pBI121-OCI was constructed, with OC-I gene under the control of CaMV35S promoter and Nos terminator.
LBA4404 strain containing this plasmid was used in genetic transformation of cabbage.
After 3d preculture, the explants were inoculated with Agrobacterium and co-cultivated for 4d at 25°C. When a slight development of the bacteria was visible on the wound of the explants, they were then transferred to regeneration medium supplied with 500 mgl-1 carbanicilin, and cultured for 7 days to kill the Agrobacterium. To select for transferred cells, explants were transferred to the same medium supplemented with 5 mgl-1 kanamycin for 6-8 weeks.
During the selection, the explants were transferred to fresh medium every 2 weeks.
The putative transformants were assayed by PCR and Southern blot analysis.
The result showed that OC-I gene was transferred into cabbage successfully.
The resistance of transgenic plants to insects was stronger compared to the control in the field.
LBA4404 strain containing this plasmid was used in genetic transformation of cabbage.
After 3d preculture, the explants were inoculated with Agrobacterium and co-cultivated for 4d at 25°C. When a slight development of the bacteria was visible on the wound of the explants, they were then transferred to regeneration medium supplied with 500 mgl-1 carbanicilin, and cultured for 7 days to kill the Agrobacterium. To select for transferred cells, explants were transferred to the same medium supplemented with 5 mgl-1 kanamycin for 6-8 weeks.
During the selection, the explants were transferred to fresh medium every 2 weeks.
The putative transformants were assayed by PCR and Southern blot analysis.
The result showed that OC-I gene was transferred into cabbage successfully.
The resistance of transgenic plants to insects was stronger compared to the control in the field.
Authors
J.J. Lei, W.J. Yang, S.H. Yuan, F.Y. Ying, L.C. Qiong
Keywords
Brassica oleracea var. capitata, cysteine proteinase inhibitor gene, transformation, Agrobacterium tumefaciens
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