Improvement in pHERD vectors to express recombinant proteins tagged with hexahistidine at either the NH₂- or COOH- terminal in Pseudomonas aeruginosa

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  • Iiyama Kazuhiro
    Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture Graduate School, Kyushu University
  • Man Lee Jae
    Laboratory of Silkworm Science, Faculty of Agriculture, Graduate School, Kyushu University
  • Kusakabe Takahiro
    Laboratory of Silkworm Science, Faculty of Agriculture, Graduate School, Kyushu University
  • Yasunaga-Aoki Chisa
    Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture Graduate School, Kyushu University
  • Shimizu Susumu
    Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture Graduate School, Kyushu University

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  • Improvement in pHERD vectors to express recombinant proteins tagged with hexahistidine at either the NH<sub>2</sub>- or COOH- terminal in <i>Pseudomonas aeruginosa</i>

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We constructed pHERDHis vectors to express polyhistidine-tagged proteins based on the pHERD vectors. The oligonucleotides containing the hexahistidine sequence were designed to be able to introduce the tag at either the NH2- or COOH- terminal. To demonstrate the usefulness of the vectors, a pyocyanin biosynthetic protein, PhzS, was expressed in a pyocyanin-deficient mutant of Pseudomonas aeruginosa. In the mutant strain, both HisPhzS (tag at the NH2- terminal) and PhzSHis (tag at the COOH- terminal) worked as a functional protein and pyocyanin was produced. It was also demonstrated that HisPhzS was purified by standard affinity chromatography.<br>

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