Agrobacterium-mediated Transformation of Human Pro-opiomelanocortin (POMC) Gene in Carrot, Daucus carota L.

• Main Authors: Rohan Noland McDonald, 麥洛翰
• Other Authors: Fure-Chyi Chen
• Format: Others
• Language: en_US
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/69174983689784233043

碩士 === 國立屏東科技大學 === 熱帶農業暨國際合作研究所 === 92 === Modern biotechnology holds great promise for improving human health as it provides relatively inexpensive production of large quantities of recombinant protein in transgenic plants. The Pro-opiomelanocortin (POMC) gene is a polypeptide precursor involved in many different important functions of the body. The objective of this experiment was to transfer the POMC gene to carrot via Agrobacterium-mediated transformation. The POMC gene constructed in a binary vector plasmid pCAMBIA 1305.1 was used to transform somatic embryos, petioles and suspension cells of carrot, Daucus carota L. Hygromycin was used to screen for transformed plants and the β-glucuronidase (GUSPlus) as a reporter gene. Concentrations of 15 mg/L, 10 mg/L and 5 mg/L were used in the selective media for somatic embryos, petioles and suspension cells, respectively. Somatic embryos proved to be the most efficient target for transformation because of greater GUS activity and post transformation regenerative capacity. The integration of the T-DNA of pCAMBIA 1305.1 into the genome of somatic embryos was confirmed by PCR, RT-PCR and Southern blot analyses.