Publication detail

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

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"
}