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Purification of the NusB gene product of Escherichia coli K12
Authors:Toshio Maekawa  Takahiro Nagase  Fumio Imamoto and Shunsuke Ishii
Institution:(1) The Institute of Medical Science, The University of Tokyo, P.O. Takanawa, 108 Tokyo, Japan;(2) The Department of Pharmacy, Science University of Tokyo, Tokyo, Japan;(3) Laboratory of Molecular Genetics, RIKEN (The Institute of Physical and Chemical Research), Hirosawa 2-1, 351-01 Wako-shi, Saitama, Japan
Abstract:Summary The nucleotide sequence of the entire nusB gene of Escherichia coli has recently been determined and the amino acid sequence of its product deduced (Ishii et al. 1984; Swindle et al. 1984). The NusB protein was purified by chromatography on Sephadex G-100, phosphocellulose and hydroxylapatite. Purification of the protein was monitored using 14C-labelled NusB protein, which was synthesized in a maxicell containing an nusB plasmid as a marker. The final product, which was at least 95% pure as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, had a molecular weight of about 16,000 and an isoelectric point of about 7.3. Analytical data on the amino acid composition of the purified protein agreed with that deduced from the DNA sequence and indicated that this protein was indeed the product of the nusB gene.Abbreviations SDS sodium dodecyl sulphate - kDa kilodaltons - bp base pair(s) - kbp kilobase pair(s)
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