Molecular cloning and expression of a mouse alpha-1,3 fucosyltransferase gene that shows homology with the human alpha-1,3 fucosyltransferase IV gene.
Molecular cloning and expression of a mouse alpha-1,3 fucosyltransferase gene that shows homology with the human alpha-1,3 fucosyltransferase IV gene.
复制标题
小鼠 α-1,3 岩藻糖基转移酶基因的分子克隆和表达，该基因与人 α-1,3 岩藻糖基转移酶 IV 基因具有同源性。
DOI：
10.1093/oxfordjournals.jbchem.a021239
复制
发表时间：
1996
影响因子：
2.7
通讯作者：
T. Muramatsu
中科院分区：
文献类型：
--
作者：
M. Ozawa;T. Muramatsu
利用聚合酶链反应从小鼠胚胎癌F9细胞中扩增并克隆了一段新的聚焦转移酶cDNA片段。用克隆的cDNA片段对小鼠基因组DNA进行Southern blot分析，发现每个单倍体基因组中有一个focusyltransferase基因拷贝。克隆了与cDNA杂交的基因组DNA的5 kb EcoRI限制性内切片段，并对包含cDNA序列的1,697 bp片段进行了测序。克隆的核苷酸序列和推导出的氨基酸序列与人α -1,3 focusyltransferase IV的同源性最高，分别为77.1和75.5%。与序列相似性一致的是，在巨细胞病毒增强子和β肌动蛋白启动子的控制下，将1544 bp的片段转染到COS-1细胞中，使细胞具有α -1,3聚焦转移酶活性，导致SSEA-1在细胞表面表达。
The polymerase chain reaction was used to amplify and clone a fragment of a novel fucosyltransferase cDNA from mouse embryonal carcinoma F9 cells. Southern blot analysis of mouse genomic DNA with the cloned cDNA fragment revealed that there is a single copy of the fucosyltransferase gene per haploid genome. A 5 kb EcoRI restriction fragment of the genomic DNA hybridized with the cDNA has been cloned and a 1,697 bp fragment containing the sequence of the cDNA was sequenced. The nucleotide sequence and the deduced amino acid sequence of the clone showed a highest degree of homology with those of human alpha-1,3 fucosyltransferase IV, i.e. 77.1 and 75.5% identity, respectively. Consistent with the sequence similarity, the transfection of a 1,544 bp fragment under the control of the cytomegarovirus enhancer and beta actin promoter into COS-1 cells confers alpha-1,3 fucosyltransferase activity on the cells, resulting in cell surface expression of SSEA-1.