Premium
Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase
Author(s) -
Ohnoutkova Ludmila,
Zitka Ondrej,
Mrizova Katarina,
Vaskova Jana,
Galuszka Petr,
Cernei Natalie,
Smedley Mark A.,
Harwood Wendy A.,
Adam Vojtech,
Kizek Rene
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200033
Subject(s) - lysine , transformation (genetics) , biology , agrobacterium , amino acid , genetically modified crops , biochemistry , biosynthesis , enzyme , gene , botany , transgene
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 C zech R epublic. 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 p B ract214::s TP dap A and p B ract214::mdap A containing the dap A gene from E scherichia 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. G olden P romise. Transgenic barley plants of the T 0 and T 1 generations were evaluated by PCR , real‐time PCR , gel electrophoresis, and Western blot. Amino acid content was analyzed by HPLC after HC l hydrolysis. The lysine content in leaves of the T 1 generation plant no. 5/5 was 50% higher than in wild‐type plants; the lysine content in seeds of T 2 generation plant no. 5/16 was 30% higher than in wild‐type seeds of spring barley cv. G olden P romise.