African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12227

Full Length Research Paper

The optimization of voltage parameter for tissue electroporation in somatic embryos of Astragalus chrysochlorus (Leguminosae)

Neslihan Turgut-Kara1* and Sule Ari1,2
1Department of Molecular Biology and Genetics, Faculty of Science, Istanbul University, 34118, Vezneciler, Istanbul, Turkey. 2Research and Application Center for Biotechnology and Genetic Engineering, 34118, Vezneciler, Istanbul, Turkey.
Email: [email protected]

  •  Accepted: 19 May 2010
  •  Published: 31 July 2010

Abstract

Somatic embryo tissues of Astragalus chrysochlorus were transformed with the b-glucuronidase (GUS) and neomycin phosphotransferase II (npt II) genes by electroporation. The effect of electric field strength was tested for transient expression. It was found that 1000 V/cm and 200 ms and 1 pulse was the optimum combination to transform embryo tissues (expression level was 61.5%). Electroporated somatic embryo tissues were positive for GUS expression and PCR analysis for the genes GUS and npt II. After PCR analysis, we found that the efficiency of the somatic embryos with transient GUS expression by electroporation was 48%.

 

Key words: Astragalus, gene transfer, electroporation, -glucuronidase, neomycin phosphotransferase II.

Abbreviation

GUS, b-glucuronidase; npt IIneomycin phosphotransferase II;nos, nopaline synthetase; IAA, indole-3-acetic acid; MS, Murashige and Skoog; 2,4-D2,4-dichloro-phenoxyacetic acid; PCR, polymerase chain reaction.