Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase

0382218 - ÚEB 2013 RIV DE eng J - Journal Article
Ohnoutková, Ludmila - Zítka, O. - Mřížová, K. - Vašková, Jana - Galuszka, P. - Cernei, N. - Smedley, M. A. - Harwood, W. A. - Adam, V. - Křížek, R.
Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase.
Electrophoresis. Roč. 33, č. 15 (2012), s. 2365-2373. ISSN 0173-0835. E-ISSN 1522-2683
R&D Projects: GA MŠMT 1M06030
Grant - others:GA MŠk(CZ) ED1.1.00/02.0068
Program: ED
Institutional research plan: CEZ:AV0Z50380511
Keywords : Foodomics * Gel electrophoresis * Polymerase chain reaction
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 3.261, year: 2012

Nutritional quality of human and animal foodstuffs is determined by the content of essential amino acids. Barley is the fourth most important cereal of the world and the second most important cereal grown in the Czech Republic. Cereal grains such as barley contain insufficient levels of some essential amino acids, especially lysine. Dihydrodipicolinate synthase is the key enzyme involved in the regulatory step for lysine biosynthesis. Two constructs pBract214::sTPdapA and pBract214::mdapA containing the dapA gene from Escherichia coli coding for the bacterial dihydrodipicolinate synthase were used for transformation of barley. An Agrobacterium-mediated technique was used for transformation of immature embryos of spring barley cv. Golden Promise. Transgenic barley plants of the T0 and T1 generations were evaluated by PCR, real-time PCR, gel electrophoresis, and Western blot. Amino acid content was analyzed by HPLC after HCl hydrolysis. The lysine content in leaves of the T1 generation plant no. 5/5 was 50% higher than in wild-type plants; the lysine content in seeds of T2 generation plant no. 5/16 was 30% higher than in wild-type seeds of spring barley cv. Golden Promise.
Permanent Link: http://hdl.handle.net/11104/0212506