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<div>This study was carried out to establish a stable genetic transformation in callus culture of Andrographis</div><div>paniculata mediated by Agrobacterium tumefaciens. The leaf disks of A. paniculata were infected with A. tumefaciens</div><div>LBA4404 carrying a binary vector pCAMBIA1304 that contain &beta;-glucuronidase (GUS) and hygromycin</div><div>phosphotransferase (hpt) genes. The infection was conducted by dipping method for one hour, followed by</div><div>co-cultivation in the dark for three days. To examine transient GUS expression, the co-cultivated leaf disks were</div><div>assayed for &beta;-glucuronidase activity and to obtain stable transformed callus, the co-cultivated leaf disks were</div><div>selected on the callus induction medium which contain 20 mg/l hygromycin for selection. The transformed</div><div>callus was periodically subcultured every three weeks into the fresh selection medium over the 15 weeks</div><div>period. To test a stable transformation, the callus was subjected to PCR analysis for GUS gene detection. The</div><div>results indicated that the co-cultivated leaf disks expressed GUS activity and proliferated to produce callus on</div><div>the selective medium. Analysis of PCR on the transformed callus indicated the presence 976 bp fragment that</div><div>confi rmed the presence of &beta;-glucuronidase gene. These fi ndings imply that the &beta;-glucuronidase was stably</div><div><br /></div><div>integrated into A. paniculata callus culture.</div><div>Keywords: Andrographis paniculata, Agrobacterium tumefaciens, andrographollide, transformed callus,</div><div>&beta;-glucuronidase gene.</div>