Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase
Language English Country Germany Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Amino Acids analysis metabolism MeSH
- Chromatography, Ion Exchange MeSH
- Hydro-Lyases biosynthesis genetics metabolism MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Plants, Genetically Modified chemistry enzymology genetics MeSH
- Hydrolysis MeSH
- Hordeum chemistry enzymology genetics MeSH
- Plant Leaves chemistry MeSH
- Lysine analysis metabolism MeSH
- Polymerase Chain Reaction MeSH
- Escherichia coli Proteins genetics metabolism MeSH
- Recombinant Proteins genetics metabolism MeSH
- Blotting, Western MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 4-hydroxy-tetrahydrodipicolinate synthase MeSH Browser
- Amino Acids MeSH
- Hydro-Lyases MeSH
- Lysine MeSH
- Escherichia coli Proteins MeSH
- Recombinant Proteins MeSH
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.
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