Original scientific paper

https://doi.org/10.5562/cca1772

Heterologous Expression of the Alba Protein from the Hyperthermophilic Archaeon Aeropyrum Pernix

Miha Črnigoj ; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia
Andrej Hanzlowsky ; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia
Tanja Vilfan ; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia
Nataša Poklar Ulrih orcid.org/0000-0002-2731-9973 ; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia; CipKeBiP – The Centre of Excellence for Integrated Approaches in Chemistry and Biology of Proteins, Jamova 39, 1000 Ljubl

Abstract

Nucleic acid binding proteins have important roles in DNA and RNA packaging, stabilisation
and repair, and in gene regulation, and they are therefore essential for all organisms. All of the known
hyperthermophiles have at least one DNA sequence encoding for the Alba proteins. The Alba proteins are
small (approximately 10 kDa), DNA-binding, basic proteins that appear to partly compensate for the lack
of histones in the archaea Aeropyrum pernix and other hyperthermophiles. Two sequences of these potential
histone counterparts, the Alba proteins, were identified in the Aeropyrum pernix genome (APE1832.1
and APE1823). By using a wide range of experimental techniques and by examining several combinations
of expression systems the expression of recombinant Alba1 and Alba2 proteins was optimized.
Co-expression of both of the Alba proteins was needed when isolating recombinant Alba2. The purification
of both recombinant Alba1 and Alba2 His-tagged proteins were simplyfied in satisfactory yield.
The electrophoretic mobility shift assay demonstrated the ability of the Alba1 and Alba2 proteins from
Aeropyrum pernix to bind DNA. (doi: 10.5562/cca1772)

Keywords

Aeropyrum pernix; hyperthermophile; archaea; Alba; DNA binding; DNA stabilisation

Hrčak ID:

75059

URI

https://hrcak.srce.hr/75059

Publication date:

20.12.2011.

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