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Expression of a staphylokinase, a thrombolytic agent in Arabidopsis thaliana

Abstract

A gene encoding staphylokinase from Staphylococcus aureus was cloned into the plant transformation binary vector pCAMBIA 1304. The transgene was introduced into the genome of A. thaliana via in planta Agrobacterium tumefaciens–mediated genetic transformation. The presence of the staphylokinase gene was confirmed by PCR in 60% of the investigated plants. The presence of the fusion protein (119 kDa) was confirmed by SDS–PAGE and Western blot analysis in protein extracts from putative transgenics. Furthermore, the amidolytic assay confirmed the activity of SAK in protein extracts in 23 out of 45 transgenic lines of A. thaliana plants.

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Abbreviations

AGL1:

Agrobacterium tumefaciens strain

bp:

Base pair

dNTP:

Deoxyribonucleotide triphosphate

EDTA:

Ethylenediaminetetraacetate

FAA:

Fixing agent (formalin, glacial acetic acid, ethanol)

gusA :

β-glucuronidase

MES:

4-Morpholineethanesulfonic acid

MS:

Murashige and skoog salts

nptII:

Neomycin phosphotransferase II

pCAMBIA:

Binary vector

PCR:

Polymerase chain reaction

PolTaq:

Polymerase from Thermococcus aquaticus

SAK:

Staphylokinase

T-DNA:

Transferred DNA

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Acknowledgments

This work was supported by The Ministry of Science and Higher Education research grant No. 6 P04B 003 18 and the University of Lodz grant No 505/040396.

Author information

Correspondence to Aneta Wiktorek-Smagur.

Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/s11274-014-1707-z.

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Wiktorek-Smagur, A., Hnatuszko-Konka, K., Geszberg, A. et al. Expression of a staphylokinase, a thrombolytic agent in Arabidopsis thaliana . World J Microbiol Biotechnol 27, 1341–1347 (2011). https://doi.org/10.1007/s11274-010-0583-4

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Keywords

  • Arabidopsis thaliana
  • Agrobacterium tumefaciens
  • Staphylokinase
  • Recombinant proteins