{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T07:15:28Z","timestamp":1791357328167,"version":"4.3.1"},"reference-count":58,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2018,7,17]],"date-time":"2018-07-17T00:00:00Z","timestamp":1531785600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2016R1E1A1A01943365"],"award-info":[{"award-number":["2016R1E1A1A01943365"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Microbiology"],"published-print":{"date-parts":[[2018,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                    is an opportunistic fungal pathogen, which causes life\u2010threatening meningoencephalitis in immunocompromised individuals and is responsible for more than 1,000,000 infections and 600,000 deaths annually worldwide. Nevertheless, anti\u2010cryptococcal therapeutic options are limited, mainly because of the similarity between fungal and human cellular structures. Owing to advances in genetic and molecular techniques and bioinformatics in the past decade,\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    , belonging to the phylum basidiomycota, is now a major pathogenic fungal model system. In particular, genetic manipulation is the first step in the identification and characterization of the function of genes for understanding the mechanisms underlying the pathogenicity of\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    . This unit describes protocols for constructing target gene deletion mutants using double\u2010joint (DJ) PCR, constitutive overexpression strains using the histone H3 gene promoter, and epitope\/fluorescence protein\u2010tagged strains in\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    . \u00a9 2018 by John Wiley &amp; Sons, Inc.\n                  <\/jats:p>","DOI":"10.1002\/cpmc.59","type":"journal-article","created":{"date-parts":[[2018,7,18]],"date-time":"2018-07-18T07:37:37Z","timestamp":1531899457000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":36,"title":["Genetic Manipulation of\n                    <i>Cryptococcus neoformans<\/i>"],"prefix":"10.1002","volume":"50","author":[{"given":"Kwang\u2010Woo","family":"Jung","sequence":"first","affiliation":[{"name":"Research Division for Biotechnology, Korea Atomic Energy Research Institute  Jeongeup Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyung\u2010Tae","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Yonsei University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yee\u2010Seul","family":"So","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Yonsei University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong\u2010Sun","family":"Bahn","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Yonsei University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,7,17]]},"reference":[{"key":"e_1_2_9_1_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00312852"},{"key":"e_1_2_9_1_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/21.14.3329"},{"key":"e_1_2_9_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2011.05.009"},{"key":"e_1_2_9_1_5_1","doi-asserted-by":"publisher","DOI":"10.1099\/00207713-50-2-931"},{"key":"e_1_2_9_1_6_1","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-18-0468"},{"key":"e_1_2_9_1_7_1","doi-asserted-by":"publisher","DOI":"10.1080\/02681219680000691"},{"issue":"8","key":"e_1_2_9_1_8_1","doi-asserted-by":"crossref","first-page":"2607","DOI":"10.1099\/00221287-148-8-2607","article-title":"A PCR\u2010based strategy to generate integrative targeting alleles with large regions of homology","volume":"148","author":"Davidson R. 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