Research on prokaryocyte expression and biological activity of the core region of Bloom's Syndrome protein
Year: 2010
Type: article
Abstract: To establish an effective approach for inducing expression of the RecQ core of Bloom’s Syndrome protein (BLM 642-1290 ) and assaying its biological activity in vitro , BLM 642-1290 recombinant protein was expressed with IPTG at room temperature in Escherichia coli , and then the expressed product was assayed using SDS-PAGE and western blotting. After purification via affinity chromatography, DNA binding activity and unwinding activity of the protein were assessed by fluorescence polarization. Furthermore, the ATPase activity of the protein was also assayed using ultraviolet spectrophotometry based on PiColorLock Gold reagent. An effective expression method was established for BLM protein in E. coli . The obvious bioactivities of the protein were observed in binding to ssDNA or dsDNA, unwinding the dsDNA in the presence of ATP, as well as catalyzing ATP hydrolysis in the presence of ssDNA in vitro . The prokaryocyte expression method of BLM 642-1290 was established successfully and the protein with biological activity was obtained from recombinant E.coli . This would be significant to provide a better understanding on BLM protein and facilitate the elucidation of mechanism of pathopoiesia in Bloom’s Syndrome. Keywords: BLM 642-1290 protein, induced expression, enzymatic activity
Source: AFRICAN JOURNAL OF BIOTECHNOLOGY
DOI: 10.5897/ajb10.633
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W2118822216