{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T12:14:17Z","timestamp":1774786457747,"version":"3.50.1"},"reference-count":22,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2010,12,23]],"date-time":"2010-12-23T00:00:00Z","timestamp":1293062400000},"content-version":"vor","delay-in-days":1421,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech and App Biochem"],"published-print":{"date-parts":[[2007,2]]},"abstract":"<jats:p>Since nonsense mutations are closely associated with severe conditions of genetic disorders, including familial cancers, rapid and precise detection of those mutations is very important for research purposes and molecular diagnosis. Currently, screening methods such as the FASAY (functional analysis of separated alleles in yeast) and the Y\u2010SC (stop codon assay in yeast) are used for functional detection of nonsense mutations in genes of interest. But these yeast\u2010based approaches are time\u2010consuming, expensive and complicated. In order to circumvent these problems, we, in the present study, devised a novel <jats:italic>Escherichia coli<\/jats:italic>\u2010based screening method, the E\u2010SC (<jats:italic>E. coli<\/jats:italic> stop codon assay) for the detection of heterozygous nonsense mutations in genes of interest. Our strategy was based on the fact that the plasmid replicating with a low copy number in <jats:italic>E. coli<\/jats:italic> allows an effective separation of normal and mutant alleles. Moreover, it relies on the expression vector, resulting in the formation of white and blue colonies for mutant and normal alleles through the expression of PCR\u2010amplified fragment\/<jats:italic>lacZ<\/jats:italic> fusion protein respectively. To evaluate the applicability of the E\u2010SC method for the detection of the heterozygous truncating mutation, PCR\u2010amplified exon 7 of the StAR [steroidogenic acute regulatory protein; causative gene of the CAH (congenital lipoid adrenal hyperplasia)] and RT (reverse transcription)\u2013PCR\u2010amplified full\u2010length cDNA of MeCP2 (methyl\u2010CpG\u2010binding protein 2; causative gene of Rett syndrome) were used. The E\u2010SC showed an almost 1:1 ratio of blue\/white colonies in all patients examined, whereas the control samples produced blue colonies only. These results demonstrate that the E\u2010SC system is useful for rapid and precise detection of known and unknown heterozygous truncation mutations in genes that cause genetic disorders and familial cancers.<\/jats:p>","DOI":"10.1042\/ba20060067","type":"journal-article","created":{"date-parts":[[2007,1,19]],"date-time":"2007-01-19T15:16:57Z","timestamp":1169219817000},"page":"137-143","source":"Crossref","is-referenced-by-count":1,"title":["Detection of heterozygous nonsense mutations in genes of interest using an <i>Escherichia coli<\/i>\u2010based stop codon assay"],"prefix":"10.1002","volume":"46","author":[{"given":"Young Joon","family":"Moon","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoonsung","family":"Kang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jee\u2010Hye","family":"Choi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kwang\u2010Ho","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2010,12,23]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/ng1093-111"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/humu.1380060402"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1385\/MB:23:2:167"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.4.1681"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.1996.0085"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1206865"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/2.10.1719"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.9.3963"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.6.2449"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1525-1578(10)60629-0"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1203795"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)62510-2"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7892608"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/13810"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(95)00419-7"},{"key":"e_1_2_6_17_2","first-page":"1.21","volume-title":"Molecular Cloning: A Laboratory Manual","author":"Sambrook J.","year":"1989"},{"key":"e_1_2_6_18_2","first-page":"13.42","volume-title":"Molecular Cloning: A Laboratory Manual","author":"Sambrook J.","year":"1989"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1177\/088307380201700108"},{"key":"e_1_2_6_20_2","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1007\/s100380300020","volume":"48","author":"Sakayori M.","year":"2003","journal-title":"J. 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