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OHNOUTKOVÁ, L. ZÍTKA, O. MRÍZOVÁ, K. VALÁŠKOVÁ, J. GALUSZKA, P. CERNEI, N. SMEDLEY, M. HARWOOD, W. ADAM, V. KIZEK, R.
Original Title
Electrophoretic and chromatographic evaluation of transgenic Barley expressing a bacterial dihydrodipicolinate synthase
Type
journal article in Web of Science
Language
English
Original Abstract
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.
Keywords
Foodomics, Gel electrophoresis, Polymerase chain reaction, Transgenic barley, Western blot
Authors
OHNOUTKOVÁ, L.; ZÍTKA, O.; MRÍZOVÁ, K.; VALÁŠKOVÁ, J.; GALUSZKA, P.; CERNEI, N.; SMEDLEY, M.; HARWOOD, W.; ADAM, V.; KIZEK, R.
RIV year
2012
Released
1. 8. 2012
ISBN
0173-0835
Periodical
Electrophoresis
Year of study
33
Number
15
State
Federal Republic of Germany
Pages from
2365
Pages to
2373
Pages count
9
BibTex
@article{BUT92847, author="Ludmila {Ohnoutková} and Ondřej {Zítka} and Katarína {Mrízová} and Jana {Valášková} and Petr {Galuszka} and Natalia Vladimirovna {Cernei} and Mark A. {Smedley} and Wendy A. {Harwood} and Vojtěch {Adam} and René {Kizek}", title="Electrophoretic and chromatographic evaluation of transgenic Barley expressing a bacterial dihydrodipicolinate synthase", journal="Electrophoresis", year="2012", volume="33", number="15", pages="2365--2373", doi="10.1002/elps.201200033", issn="0173-0835" }