{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:25:49Z","timestamp":1761611149890},"reference-count":26,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2005,2,1]],"date-time":"2005-02-01T00:00:00Z","timestamp":1107216000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2005,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            We describe a novel method of random chimeragenesis based on highly frequent deletion formation in the\n            <jats:italic>Escherichia coli ssb-3<\/jats:italic>\n            strain and a deletion-directed chimera selection system that uses the\n            <jats:italic>rpsL<\/jats:italic>\n            <jats:sup>+<\/jats:sup>\n            gene as a reporter. It enables the selection of chimeras without target gene expression and can therefore be applied to cytotoxic targets. When this system was applied to phospholipase D genes from\n            <jats:italic>Streptomyces septatus<\/jats:italic>\n            TH-2 and\n            <jats:italic>Streptomyces halstedii<\/jats:italic>\n            subsp.\n            <jats:italic>scabies<\/jats:italic>\n            K6 (examples of cytotoxic targets), chimeragenesis occurred between short identical sequences at the corresponding position of the parental genes with large variations. Chimeragenesis was &gt;1,000 times more frequent in the\n            <jats:italic>ssb-3<\/jats:italic>\n            background than in the\n            <jats:italic>ssb<\/jats:italic>\n            <jats:sup>+<\/jats:sup>\n            background. We called this system repeat-length-independent broad-spectrum shuffling. It enables the convenient chimeragenesis and functional study of chimeric proteins. In fact, we found two amino acid residues related to the thermostability of phospholipase D (Phe426 and Thr433) by comparing thermostability among the chimeric enzymes obtained.\n          <\/jats:p>","DOI":"10.1128\/aem.71.2.754-760.2005","type":"journal-article","created":{"date-parts":[[2005,2,3]],"date-time":"2005-02-03T18:24:28Z","timestamp":1107455068000},"page":"754-760","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":16,"title":["Repeat-Length-Independent Broad-Spectrum Shuffling, a Novel Method of Generating a Random Chimera Library In Vivo"],"prefix":"10.1128","volume":"71","author":[{"given":"Koichi","family":"Mori","sequence":"first","affiliation":[{"name":"Research Institute for Biological Sciences, Okayama, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takafumi","family":"Mukaihara","sequence":"additional","affiliation":[{"name":"Research Institute for Biological Sciences, Okayama, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshiko","family":"Uesugi","sequence":"additional","affiliation":[{"name":"Research Institute for Biological Sciences, Okayama, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masaki","family":"Iwabuchi","sequence":"additional","affiliation":[{"name":"Research Institute for Biological Sciences, Okayama, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tadashi","family":"Hatanaka","sequence":"additional","affiliation":[{"name":"Research Institute for Biological Sciences, Okayama, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(95)00313-U"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.81.22.6929"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.134.3.1141-1156.1978"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/34663"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/0003-2670(94)00613-Q","volume":"304","year":"1995","unstructured":"D'Arrigo, P., V. 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