{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T04:17:04Z","timestamp":1773980224628,"version":"3.50.1"},"reference-count":39,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2011,12,3]],"date-time":"2011-12-03T00:00:00Z","timestamp":1322870400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Reprod Domestic Animals"],"published-print":{"date-parts":[[2012,10]]},"abstract":"<jats:title>Contents<\/jats:title><jats:p>Porcine embryonic fibroblasts (PEFs) are widely used as donor cells for somatic cell nuclear transfer (SCNT) in pigs. Transfection of PEFs with exogenous DNA is essential for producing genetically modified (GM; transgenic or knockout) pigs via SCNT. In this case, selectable markers are strictly required selecting and enriching stably transfected cells. The most frequently used selective drug for this purpose is a neomycin analogue (G418\/geneticin); <jats:italic>neo<\/jats:italic> has been widely used as a selectable marker gene in the genomic manipulation of pigs. However, little is known about optimal concentrations of other selection drugs. This often hampers functional analysis of the porcine genome and development of individual GM pigs. This study explores the optimal concentrations of selective drugs, other than neomycin, that can be used for the selection of transfected PEFs. Porcine embryonic fibroblasts were incubated in media containing different concentrations of drugs for up to 10\u2003days, to determine the optimal drug concentrations fatal for PEFs. The following concentrations were found to be optimal selective concentrations for use with PEFs: G418\/geneticin, 400\u2003\u03bcg\/ml; blasticidin S, 8\u2003\u03bcg\/ml; hygromycin B, 40\u2003\u03bcg\/ml; puromycin, 2\u2003\u03bcg\/ml; and zeocin, 800\u2003\u03bcg\/ml. Repeated transfections with plasmids carrying selectable markers resulted in the generation of multidrug\u2010resistant swine transfectants. Furthermore, these markers were found to be independent. The present information will be useful for the production of SCNT\u2010mediated GM piglets that express multiple transgenes.<\/jats:p>","DOI":"10.1111\/j.1439-0531.2011.01964.x","type":"journal-article","created":{"date-parts":[[2011,12,3]],"date-time":"2011-12-03T22:23:38Z","timestamp":1322951018000},"page":"759-765","source":"Crossref","is-referenced-by-count":19,"title":["Determination of the Optimal Concentration of Several Selective Drugs Useful for Generating Multi\u2010Transgenic Porcine Embryonic Fibroblasts"],"prefix":"10.1111","volume":"47","author":[{"given":"M","family":"Sato","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M","family":"Ohtsuka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H","family":"Miura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K","family":"Miyoshi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S","family":"Watanabe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2011,12,3]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-3089.2008.00457.x"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/3.9.2303"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02401419"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1089\/clo.2008.0036"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1080\/10495390903048235"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.125.1.145"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm0995-964"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0302-251"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0019986"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-4-16"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19980105)57:1<1::AID-BIT1>3.0.CO;2-M"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0598-468"},{"key":"e_1_2_8_14_1","volume-title":"DNA Cloning: A Practical Approach Volume 4: Mammalian Systems (Practical Approach Series)","author":"Glover M","year":"2011"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1089\/clo.2004.6.327"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4827(91)90427-V"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb1961.51.3165"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb1961.55.3155"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0307819101"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(200006)56:2<145::AID-MRD4>3.0.CO;2-8"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/1477-7827-1-82"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1068228"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1198"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.11.6.3374"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.5938\/youton.28.211"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1089\/clo.2007.0021"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(91)90434-D"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11427-010-0080-x"},{"key":"e_1_2_8_29_1","first-page":"1268","article-title":"Hygromycin I. 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