{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T00:55:08Z","timestamp":1793753708932,"version":"4.6.1"},"reference-count":28,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2012,8,1]],"date-time":"2012-08-01T00:00:00Z","timestamp":1343779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2012,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>The Red recombinase system, the most convenient genetic tool applied in Escherichia coli and other bacteria, was introduced for gene replacement in Klebsiella pneumoniae. The novel K. pneumoniae gene replacement system comprised the Red and FLP recombinases expression vector pDK6-red and pDK6-flp, and linear DNA fragments which encompassed a selective marker gene with target gene flanking extensions; the latter were PCR amplified using a plasmid DNA template obtained by in vivo recombination in E. coli. In this study, dhak1 gene, encoding a subunit of dihydroxyacetone kinase II, was deleted markerlessly at a transformation ratio of 260 CFU\/\u03bcg DNA, i.e., 1,000-fold higher than that achieved in the native way. Our studies provide an efficient method with detailed protocol to perform gene replacement in K. pneumoniae and has great potential to be developed as a routine genetic approach for this important industrial microorganism.<\/jats:p>","DOI":"10.1007\/s10295-012-1117-x","type":"journal-article","created":{"date-parts":[[2012,3,19]],"date-time":"2012-03-19T15:10:39Z","timestamp":1332169839000},"page":"1219-1226","source":"Crossref","is-referenced-by-count":58,"title":["Red recombinase assisted gene replacement in <i>Klebsiella pneumoniae<\/i>"],"prefix":"10.1093","volume":"39","author":[{"given":"Dong","family":"Wei","sequence":"first","affiliation":[{"name":"grid.413109.e 0000000097356249 Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science & Technology), Ministry of Education, College of Biotechnology Tianjin University of Science and Technology 300457 Tianjin People\u2019s Republic of China"},{"name":"grid.9227.e 0000000119573309 Lab of Biorefinery, Shanghai Advanced Research Institute Chinese Academy of Sciences No. 99 Haike Road 201210 Pudong, Shanghai People\u2019s Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Wang","sequence":"additional","affiliation":[{"name":"grid.413109.e 0000000097356249 Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science & Technology), Ministry of Education, College of Biotechnology Tianjin University of Science and Technology 300457 Tianjin People\u2019s Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiping","family":"Shi","sequence":"additional","affiliation":[{"name":"grid.9227.e 0000000119573309 Lab of Biorefinery, Shanghai Advanced Research Institute Chinese Academy of Sciences No. 99 Haike Road 201210 Pudong, Shanghai People\u2019s Republic of 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