小鼠α 1,2-聚焦转移酶基因的克隆与表达

Shi yan sheng wu xue bao Pub Date : 2005-12-01
Bei Lin, Makiko Saito, Masao Iwamori
{"title":"小鼠α 1,2-聚焦转移酶基因的克隆与表达","authors":"Bei Lin,&nbsp;Makiko Saito,&nbsp;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,&nbsp;Makiko Saito,&nbsp;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}
引用次数: 0

摘要

利用RT-PCR技术克隆了小鼠α 1,2- focusyltransferase的DNA片段MFUT-II。将开放阅读框连接到哺乳动物表达载体pcDNA3.1 (pcDNA3.1-MFUT-II)上,使用Cellphect转染试剂盒瞬时转染COS-7细胞。此外,通过Northern杂交分析该基因在小鼠组织中的表达,在多个组织中显示出3.5 kb的mRNA转录产物。采用Southern blot检测MFUT-II的存在。MFUT-II是小鼠α 1,2- ft基因家族的新成员。其开放阅读框编码347个氨基酸,预测分子量为39.21 kDa,与小鼠H和Sec1基因序列同源。MFUT-II与人Se基因(79.0%)、大鼠FTB基因(89.0%)和兔FT-III基因(77.0%)序列同源。pcDNA3.1-MFUT- II转染的COS-7细胞显示α 1,2-聚焦转移酶活性。Southern分析显示,MFUT-II作为单拷贝基因存在于小鼠基因组中。总之,MFUT-II是一个小鼠硒基因。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信