کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1952527 1057213 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein
چکیده انگلیسی

HU is a most abundant DNA-binding protein in bacteria. This protein is conserved either in its heterodimeric form or in one of its homodimeric forms in all bacteria, in plant chloroplasts, and in some viruses. HU protein non-specifically binds and bends DNA as a hetero- or homodimer and can participate in DNA supercoiling and DNA condensation. It also takes part in some DNA functions such as replication, recombination, and repair. HU does not recognize any specific sequences but shows some specificity to cruciform DNA and to repair intermediates, e.g., nick, gap, bulge, 3′-overhang, etc. To understand the features of HU binding to DNA and repair intermediates, a fast and easy HU proteins purification procedure is required. Here we report overproduction and purification of the HU homodimers. The method of HU purification allows obtaining a pure recombinant non-tagged protein cloned in Escherichia coli. We applied this method for purification of Acholeplasma laidlawii HU and demonstrated that this protein possesses a DNA – binding activity and is free of contaminating nuclease activity. Besides that we have shown that expression of A. laidlawii ihf_hu gene in a slow-growing hupAB E. coli strain restores the wild-type growth indicating that aclHU can perform the basic functions of E. coli HU in vivo.


► Acholeplasma laidlawii HU protein cloned and expressed in Escherichia coli.
► Fast and easy recombinant protein non-affinity purification procedure developed.
► Absence of nuclease activity and E. coli histone-like proteins impurities were shown.
► A. laidlawii HU protein has the smallest known DNA-binding site for 8 bp.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimie - Volume 93, Issue 7, July 2011, Pages 1102–1109
نویسندگان
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