{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T18:40:03Z","timestamp":1769020803362,"version":"3.49.0"},"reference-count":24,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2003,3,13]],"date-time":"2003-03-13T00:00:00Z","timestamp":1047513600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[2003,4]]},"abstract":"<jats:p>The transcription patterns of 64 linear double stranded DNA templates obtained with T7 RNA polymerase were investigated. These templates consisted of 17\u2003nucleotide\u2010long sequences under the control of the minimal bacteriophage T7 promoter and represented all possible combinations of nucleotides at positions +8, +10 and +11. Two clearly distinct types of template were identified, which produced the range of transcription patterns observed: (a) those that yielded 17\u2010nucleotide\u2010long RNA as the only detectable run\u2010off product (only 15% of the total), and (b) templates that in addition to the expected full\u2010length RNA, produced other products longer than 17 nucleotides. Self\u2010complementarity analysis of the expected run\u2010off transcripts showed that those obtained from the first type of template were able to form stable intermolecular duplexes with non\u2010base\u2010paired 3\u2032\u2010ends. However, the second type of template yielded RNAs able to generate energetically favorable intermolecular duplexes with 3\u2032\u2010end complementarity, therefore yielding an RNA\u2010primed RNA\u2010template. The gel\u2010purified 17\u2010nucleotide\u2010long RNAs transcribed from the latter yielded longer products when incubated under <jats:italic>in\u2003vitro<\/jats:italic> transcription conditions in the absence of a DNA template. No extension was observed when assaying the 17\u2010nucleotide RNA products resulting from the first type of template. We observed that just a single nucleotide change within the DNA template could convert the RNA product from an RNA\u2010primed template into a nonextendible dimer thus leading to a drastic switch of the 17\u2010nucleotide product yield from less than 10% to 100%. Further, two type B DNA templates were extended by two nucleotides at the 3\u2032\u2010end, to produce RNA transcripts theoretically unable to form 3\u2032\u2010end base\u2010paired duplexes. The full\u2010length products of these modified DNA templates were found to be nonextendible by T7 RNA polymerase under the standard <jats:italic>in vitro<\/jats:italic> transcription conditions.<\/jats:p>","DOI":"10.1046\/j.1432-1033.2003.03510.x","type":"journal-article","created":{"date-parts":[[2004,12,23]],"date-time":"2004-12-23T02:52:42Z","timestamp":1103770362000},"page":"1458-1465","source":"Crossref","is-referenced-by-count":57,"title":["Preventing nondesired RNA\u2010primed RNA extension catalyzed by T7 RNA polymerase"],"prefix":"10.1111","volume":"270","author":[{"given":"Genoveva A.","family":"Nacheva","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alfredo","family":"Berzal\u2010Herranz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2003,3,17]]},"reference":[{"key":"e_1_2_16_2_2","first-page":"(","volume-title":"The Enzymes","author":"Chamberlin M.","year":"1982"},{"key":"e_1_2_16_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(92)90997-X"},{"key":"e_1_2_16_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi9630467"},{"key":"e_1_2_16_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.286.5448.2305"},{"key":"e_1_2_16_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2000.4150"},{"key":"e_1_2_16_7_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2000.4114"},{"key":"e_1_2_16_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/15.21.8783"},{"key":"e_1_2_16_9_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/26.15.3550"},{"key":"e_1_2_16_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(89)90917-3"},{"key":"e_1_2_16_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(90)90456-O"},{"key":"e_1_2_16_12_2","doi-asserted-by":"crossref","first-page":"3458","DOI":"10.1002\/j.1460-2075.1996.tb00712.x","article-title":"Template\u2010free generation of RNA species that replicate with bacteriophage T7 RNA polymerase","volume":"15","author":"Bierbricher C.K.","year":"1996","journal-title":"EMBO J."},{"key":"e_1_2_16_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.15.6972"},{"key":"e_1_2_16_14_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.11.6298"},{"key":"e_1_2_16_15_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/17.8.3023"},{"key":"e_1_2_16_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00167a044"},{"key":"e_1_2_16_17_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.81.7.2035"},{"key":"e_1_2_16_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/15.6.2653"},{"key":"e_1_2_16_19_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.41.29376"},{"key":"e_1_2_16_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(89)80091-6"},{"key":"e_1_2_16_21_2","doi-asserted-by":"publisher","DOI":"10.1089\/oli.1.1999.9.33"},{"key":"e_1_2_16_22_2","doi-asserted-by":"publisher","DOI":"10.1089\/oli.1.1999.9.433"},{"key":"e_1_2_16_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00411a012"},{"key":"e_1_2_16_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00106a036"},{"key":"e_1_2_16_25_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/30.10.e46"}],"container-title":["European Journal of Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1046%2Fj.1432-1033.2003.03510.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1046\/j.1432-1033.2003.03510.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T13:30:58Z","timestamp":1697463058000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1046\/j.1432-1033.2003.03510.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,3,17]]},"references-count":24,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2003,4]]}},"alternative-id":["10.1046\/j.1432-1033.2003.03510.x"],"URL":"https:\/\/doi.org\/10.1046\/j.1432-1033.2003.03510.x","archive":["Portico"],"relation":{},"ISSN":["0014-2956","1432-1033"],"issn-type":[{"value":"0014-2956","type":"print"},{"value":"1432-1033","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,3,17]]}}}