Transgenic IR64 rice seeds with increased lysine content
2011
Oliva, N. | Abrigo, E. | Balindong, J. | Trijatmiko, K. | Dueñas, O. | Chadha-Mohanty, P. | Barry, G. | Slamet-Loedin, I., International Rice Research Inst., DAPO Box 7777, Manila (Philippines). Genetic Transformation Lab.
Rice is deficient in essential amino acids, with lysine being the most limiting amino acid for dietary requirements. Improving quality of protein in rice can be a cost-effective way to combat the protein-calorie malnutrition in Asia. It has previously been shown that expression of Corynobacterium gluatamicum dehydrodipicolinate synthase(DHPS),an enzyme regulating the lysine biosynthetic pathway,increase the free lysine content of canola and soybean seeds.Herein, transformation of expression vectors using the C glutamicum dapA.gene expressing DHPS under the control of rice endosperm specific promoters,glutelin B-1 and glutelin B-4 (p C1300 int-GluB1p-mcts dapA and pC1300 int-GluB4p-mcts-dapA) were made with IR64 as the recipient donor plant. Four hundred eight transformants were obtained from both constructs. Forty eight single copy plants were determined using Southern blot analysis. ELISA results in T1 and T2 generation show that there is a 2-5 fold increase in expression levels of cap A protein.Free lysine contents of seeds of a transgenic event using High Performance Liquid Chromatography(HPLC)was shown to be 43 ug/g compared to 4.5 ug/g for wild type IR64.
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