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| Cloning of Soybean Oleosin and EGF Fusion Gene and Expression in Safflower Seeds |
| PANG Shi-feng1, JIANG Chao2, LI Wen-rong1,2, FENG Zhi-guo2, LIU Min2, CHU Sheng-hui2, LI Xiao-kun2, ZHENG Ke-qin1 |
1. Guangdong Medical College, Dongguang 523808, China;
2. College of Pharmacy, Wenzhou Medical University, Wenzhou 325035, China |
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Abstract Aim: The purpose is expressing EGF in oil-body of transgenic safflower. Method: The Ddoil-EGF fusion gene was got with polymerase chain reaction (PCR) amplification, and the Ddoil-EGF fusion gene was cloned into plant expression vector pCAMBIA1390R to gain p1390Do-EGF. The recombinant plasmid p1390Do-EGF was transferred into Agrobacterium tumefaciens LBA4404. By using co-cultivated method,the fusion gene Ddoil-EGF was transferred into safflower cells. Transferred shoots were selected on solid medium containing mannose.EGF was detected by PCR and real-time RT-PCR in safflower. The expression of EGF in safflower was detected by SDS-PAGE and Western blot. Effect of EGF on the proliferation of Bab/c 3T3 cells assessed by MTT. Result: PCR result showed that EGF gene was transformed into safflower. Relative quantitative real-time RT-PCR showed EGF mRNA was transcripted in safflower. The transgenic safflower seeds expressing Ddoil-EGF fusion protein through SDS-PAGE were confirmed. Western blot result showed that recombinant protein expressed by transgenic safflower could eliciting immunoresponse to EGF antibody. MTT result showed that EGF can stimulate the proliferation of Bab/c 3T3 cells. Conclusion:The Ddoil-EGF fusion gene was transformed into safflower and was expressed in safflower seeds.It is a new ways to express EGF in plant.
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Received: 20 January 2014
Published: 25 April 2014
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