Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase
Jazyk angličtina Země Německo Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
22887157
DOI
10.1002/elps.201200033
Knihovny.cz E-zdroje
- MeSH
- aminokyseliny analýza metabolismus MeSH
- chromatografie iontoměničová MeSH
- dehydratasy biosyntéza genetika metabolismus MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- geneticky modifikované rostliny chemie enzymologie genetika MeSH
- hydrolýza MeSH
- ječmen (rod) chemie enzymologie genetika MeSH
- listy rostlin chemie MeSH
- lysin analýza metabolismus MeSH
- polymerázová řetězová reakce MeSH
- proteiny z Escherichia coli genetika metabolismus MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- western blotting MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 4-hydroxy-tetrahydrodipicolinate synthase MeSH Prohlížeč
- aminokyseliny MeSH
- dehydratasy MeSH
- lysin MeSH
- proteiny z Escherichia coli MeSH
- rekombinantní proteiny 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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