RRC ID 36336
著者 Iiyama K, Lee JM, Tatsuke T, Mon H, Kusakabe T.
タイトル Expression and Characterization of Recombinant Serratia liquefaciens Nucleases Produced with Baculovirus-mediated Silkworm Expression System.
ジャーナル Mol Biotechnol
Abstract Baculovirus-Bombyx mori protein expression system has mainly been used for translation of eukaryotic proteins. In contrast, information pertaining to bacterial protein expression using this system is not sufficient. Therefore, recombinant nucleases from Serratia liquefaciens (rSlNucAs) were expressed in a Baculovirus-B. mori protein expression system. rSlNucAs containing the native signal peptide (rSlNucA-NSP) or silkworm 30-K signal peptide (rSlNucA-30K) at the NH2-terminus were constructed to enable secretion into the extracellular fraction. Both rSlNucA-30K and rSlNucA-NSP were successfully secreted into hemolymph of B. mori larvae. Affinity-purified rSlNucAs showed high nuclease activity. Optimum pH was 7.5 and half of maximum activity was maintained between pH 7.0 and 9.5. Optimum temperature was 35 °C. rSlNucAs showed sufficient activity in twofold-diluted radioimmunoprecipitation assay buffer and undiluted, mild lysis buffer. Genomic DNA of Escherichia coli was efficiently digested by rSlNucAs in the bacterial lysate. The results in this study suggest that rSlNucAs expressed by the Baculovirus-B. mori protein expression system will be a useful tool in molecular biology. Functional recombinant protein of bacteria was produced by Baculovirus-B. mori protein expression system. This system may be highly suitable for bacterial extracellular protein secreted via Sec pathway.
巻・号 58(6)
ページ 393-403
公開日 2016-6-1
DOI 10.1007/s12033-016-9937-y
PII 10.1007/s12033-016-9937-y
PMID 27059494
MeSH Animals Bacterial Proteins / genetics Bacterial Proteins / metabolism Baculoviridae / enzymology Baculoviridae / genetics* Bombyx / virology* Cloning, Molecular Deoxyribonucleases / chemistry Deoxyribonucleases / genetics* Deoxyribonucleases / metabolism* Gene Expression Hydrogen-Ion Concentration Recombinant Proteins / chemistry Recombinant Proteins / metabolism Serratia liquefaciens / enzymology* Serratia liquefaciens / genetics Temperature
IF 2.022
引用数 1
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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