Studies on Transformation of Functional Peptide VVYP Genes with Hypotriglyceridemic Action into Transplastomic Rice

• Main Authors: Jia-Yu Lee, 李家宇
• Other Authors: 曾夢蛟
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/97268878365195219545

碩士 === 國立中興大學 === 園藝學系所 === 103 === Rice(Oryza sativa L.) is one of the most important crops grown worldwide, and also is the staple food and vital crop for the inhabitants in Taiwan. Currently, gene transformation offers new possibilities for obtaining high yielding, better quality rice cultivars, and also those resistant to major diseases. Using transgenic crops as bioreactors to produce industrial, feed and fodder additives, and pharmaceutical proteins become a new way for increasing economic values of crops and may solve the problems we are facing. Tetra-peptide, VVYP, is known to inhibit fat absorption in the digestive tract and to enhance the activity of hepatic triglyceride lipase to increase the clearance of body fat. Expression of foreign genes via chloroplast genomes offers several unique advantages, including high protein levels, expressing multiple genes in operons, transgene containment by maternal inheritance, as well as lack of gene silencing, position and pleiotropic effects. In this study, Gy5-9VVYP, βC-7VVYP and15VVYP genes are transformed into therice chloroplast by using daao/aadA gene as selectable marker gene. The objective of this study is to develop the rice chloroplast as a bioreactor for production of functional VVYP peptide with hypotriglyceridemic action. Chloroplast transformation vectors harboring the VVYP and daao/aadA genes had been constructed and transferred into the rice chloroplast via biolistic bombardment. The regenerated transplastomic plantlets were selected by D-alanine and spectinomycin. The results of PCR and RT-PCR analysis of T0 and T1 leaves indicated that the transformed genes are present in the chloroplast genome of transplastomic rice plants, and expressed its mRNA.