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

被引:2
|
作者
Crnigoj, Miha [1 ]
Hanzlowsky, Andrej [1 ]
Vilfan, Tanja [1 ]
Ulrih, Natasa Poklar [1 ,2 ]
机构
[1] Univ Ljubljana, Biotech Fac, Dept Food Sci & Technol, Ljubljana 1000, Slovenia
[2] CipKeBiP Ctr Excellence Integrated Approaches Che, Ljubljana 1000, Slovenia
关键词
Aeropyrum pernix; hyperthermophile; archaea; Alba; DNA binding; DNA stabilisation; DNA-BINDING PROTEIN; X-RAY ANALYSIS; CHROMATIN PROTEIN; CRYSTALLIZATION;
D O I
10.5562/cca1772
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
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 APE 1823). By using a wide range of experimental techniques and by examining several combinations of expression systems the expression of recombinant Alba 1 and Alba2 proteins was optimized. Co-expression of both of the Alba proteins was needed when isolating recombinant Alba2. The purification of both recombinant Alba 1 and Alba2 His-tagged proteins were simplyfied in satisfactory yield. The electrophoretic mobility shift assay demonstrated the ability of the Alba 1 and Alba2 proteins from Aeropyrum pernix to bind DNA. (doi: 10.5562/cca1772)
引用
收藏
页码:499 / 504
页数:6
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