Constructing of Sense and Antisense pepc Expression Vector of Jatropha curcas and the Transformation

FAN Zheng-qi; LU Meng-zhu; LI Ji-yuan; TIAN Min; LI Xin-lei; WU Kai-yun; CHEN Dong-liang; FAN Miao-hua

Antisense pepc gene expression vector was constructed by the 597 bp sequence of pepc gene ligated to pBI121 in the antisense orientation. And sense expression vector was recombined by the full-length (3 000 bp) sequence inserted to pCAMBIA2300. Mediated by Agrobacterium tumefaciens, the sense and antisense genes were transferred into the tobacco using leaf-disk method. Results of PCR and PCR-Southern determination indicated two gene sequences were integrated into the tobacco genome in the sense and antisense orientation. The activity of PEPCase was determined. It was found that the fat content of transgenic tobacco with sense pepc was 44.3 percent lower than that of control, and that with antisense pepc was 26.3 percent higher than that of the control.

1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China

2. Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Received Date: 2010-03-19

Abstract: Antisense pepc gene expression vector was constructed by the 597 bp sequence of pepc gene ligated to pBI121 in the antisense orientation. And sense expression vector was recombined by the full-length (3 000 bp) sequence inserted to pCAMBIA2300. Mediated by Agrobacterium tumefaciens, the sense and antisense genes were transferred into the tobacco using leaf-disk method. Results of PCR and PCR-Southern determination indicated two gene sequences were integrated into the tobacco genome in the sense and antisense orientation. The activity of PEPCase was determined. It was found that the fat content of transgenic tobacco with sense pepc was 44.3 percent lower than that of control, and that with antisense pepc was 26.3 percent higher than that of the control.

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Reference (21)

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Constructing of Sense and Antisense pepc Expression Vector of Jatropha curcas and the Transformation

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