The expression of analgesic-antitumor peptide (AGAP) from Chinese Buthus martensii Karsch in transgenic tobacco and tomato.
The present study aimed to obtain analgesic-antitumor peptide (AGAP) gene expression in plants. The analgesic-antitumor peptide (AGAP) gene was from the venom of Buthus martensii Karsch. Previous studies showed that AGAP has both analgesic and antitumor activities, suggesting that AGAP would be useful in clinical situations as an antitumor drug. Given that using a plant as an expression vector has more advantages than prokaryotic expression, we tried to obtain transgenic plants containing AGAP. In the present study, the AGAP gene was cloned into the plasmid pBI121 to obtain the plant expression vector pBI-AGAP. By tri-parental mating and freeze-thaw transformation, pBI-AGAP was transformed into Agrobacterium tumefaciens LBA4404. Tobacco (Nicotiana tabacum) and tomato (Lycopersicom esculentum) were transformed by the method of Agrobacterium-mediated leaf disc transformation. The transformants were then screened to grow and root on media containing kanamycin. Finally, transformations were confirmed by analysis of PCR, RT-PCR and western blotting. The results showed that the AGAP gene was integrated into the genomic DNA of tobacco and tomato and was successfully expressed. Therefore, the present study suggests a potential industrial application of AGAP expressed in plants.
School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning Province, People's Republic of China.
Linlin
L
Lai
Tingting
T
Huang
Yi
Y
Wang
Yanfeng
Y
Liu
Jinghai
J
Zhang
Yongbo
Y
Song
D016960:Agrobacterium tumefaciens; D015153:Blotting, Western; D002681:China; D005822:Genetic Vectors / Q000235:genetics; D018551:Solanum lycopersicum / Q000235:genetics*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000378:metabolism; D012038:Regeneration; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012604:Scorpion Venoms / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic
D011994:Recombinant Proteins; D012604:Scorpion Venoms; C477804:antitumor-analgesic peptide, Buthus martensii