Daum

Abstract
Introduction
Materials and Methods
Results and Discussion
Acknowledgements
Literature Cited

  The amorpha-4, 11-diene synthase gene isolated from Artemisia annua was inserted into the binary vector pILTAB 357 within restriction enzyme sites, EcoRI and BamHI. It was introduced into the lettuce through Agrobacterium-mediated transformation. A total of nine transgenic plants were obtained from the inoculated lettuce leaf disc. The integration of the amorpha-4, diene synthase gene into the lettuce genome was confirmed by polymerase chain reaction (PCR) and Southern hybridization. The PCR products of 1.6 kb have been produced from all transgenic plants. From the Southern blot analysis, the amorpha-4, 11-diene synthase gene was transferred to nine transgenic lettuce plants with one to three copies. The antibiotics resistance of T1 transgenic plants was tested by growing on medium supplemented with kanamycin sulfate (0, 50, and 100 ㎎ㆍL?¹). In the result, the segregation ratio of T1 transgenic plants was exhibited about 2.7:1, 638 individuals of kanamycin-resistant and 236 of kanamycin-sensitive. The inheritance characteristics of amorpha-4, 11-diene synthase gene in the T1 transgenic plants was analyzed by PCR. The 1.6 kb PCR products were produced from nine lines of T1 transgenic plants and its segregation ratio was 1:1. The artemisinin content of T1 transgenic lettuces was analyzed through gas chromatography and mass spectrometry. The gas chromatography analysis showed that the multi-peaks were produced from nine lines of T1 transgenic plants at a retention time of 10 min 7 s. Thus, mass spectrometric analysis of the T1 transgenic plants was performed. In the result, however, no T1 transgenic plants produce any peak corresponding to artemisinin.