{"title":"小鼠α 1,2-聚焦转移酶基因的克隆与表达","authors":"Bei Lin, Makiko Saito, Masao Iwamori","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>MFUT-II, a DNA fragment encoding murine alpha1,2-fucosyltransferase,was cloned by RT-PCR. The open-reading frame was ligated into mammalian expression vector pcDNA 3.1 (pcDNA3.1-MFUT-II) and was transiently transfected into COS-7 cells using a Cellphect Transfection kit. Moreover, the expression of the gene in murine tissues was analyzed with Northern hybridization showing a 3.5-Kb mRNA transcript product in several tissues. The existence of MFUT-II was detected by Southern blot. MFUT-II was found to be a new member of the murine alpha1,2-FT gene family. Its open reading frame encoded 347 amino acids with a predicted molecular mass of 39.21 kDa and exhibited sequence homology with murine H & Sec1 gene. MFUT-II also exhibited sequence homology with Human Se gene (79.0%), Rat FTB gene (89.0%) and Rabbit FT-III gene (77.0%), respectively. COS-7 cells transfected with pcDNA3.1-MFUT- II showed alpha 1,2-fucosyltransferase activity. Southern analysis revealed that MFUT-II was present in the mouse genome as a single-copy gene. In conclusion, MFUT-II was a murine Se gene.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 6","pages":"536-44"},"PeriodicalIF":0.0000,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Cloning and expression of murine alpha1,2-fucosyltransferase gene].\",\"authors\":\"Bei Lin, Makiko Saito, Masao Iwamori\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>MFUT-II, a DNA fragment encoding murine alpha1,2-fucosyltransferase,was cloned by RT-PCR. The open-reading frame was ligated into mammalian expression vector pcDNA 3.1 (pcDNA3.1-MFUT-II) and was transiently transfected into COS-7 cells using a Cellphect Transfection kit. Moreover, the expression of the gene in murine tissues was analyzed with Northern hybridization showing a 3.5-Kb mRNA transcript product in several tissues. The existence of MFUT-II was detected by Southern blot. MFUT-II was found to be a new member of the murine alpha1,2-FT gene family. Its open reading frame encoded 347 amino acids with a predicted molecular mass of 39.21 kDa and exhibited sequence homology with murine H & Sec1 gene. MFUT-II also exhibited sequence homology with Human Se gene (79.0%), Rat FTB gene (89.0%) and Rabbit FT-III gene (77.0%), respectively. COS-7 cells transfected with pcDNA3.1-MFUT- II showed alpha 1,2-fucosyltransferase activity. Southern analysis revealed that MFUT-II was present in the mouse genome as a single-copy gene. In conclusion, MFUT-II was a murine Se gene.</p>\",\"PeriodicalId\":77395,\"journal\":{\"name\":\"Shi yan sheng wu xue bao\",\"volume\":\"38 6\",\"pages\":\"536-44\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2005-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Shi yan sheng wu xue bao\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Shi yan sheng wu xue bao","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
[Cloning and expression of murine alpha1,2-fucosyltransferase gene].
MFUT-II, a DNA fragment encoding murine alpha1,2-fucosyltransferase,was cloned by RT-PCR. The open-reading frame was ligated into mammalian expression vector pcDNA 3.1 (pcDNA3.1-MFUT-II) and was transiently transfected into COS-7 cells using a Cellphect Transfection kit. Moreover, the expression of the gene in murine tissues was analyzed with Northern hybridization showing a 3.5-Kb mRNA transcript product in several tissues. The existence of MFUT-II was detected by Southern blot. MFUT-II was found to be a new member of the murine alpha1,2-FT gene family. Its open reading frame encoded 347 amino acids with a predicted molecular mass of 39.21 kDa and exhibited sequence homology with murine H & Sec1 gene. MFUT-II also exhibited sequence homology with Human Se gene (79.0%), Rat FTB gene (89.0%) and Rabbit FT-III gene (77.0%), respectively. COS-7 cells transfected with pcDNA3.1-MFUT- II showed alpha 1,2-fucosyltransferase activity. Southern analysis revealed that MFUT-II was present in the mouse genome as a single-copy gene. In conclusion, MFUT-II was a murine Se gene.