申请人:The Regents of the University of Michigan
公开号:US05595900A1
公开(公告)日:1997-01-21
A method for isolating a gene, comprising: (i) isolating a cell possessing a post-translational characteristic of interest, said post-translational characteristic being the presence of a membrane-bound oligosaccharide or polysaccharide of interest on the surface of said cell, the presence of a soluble oligosaccharide or polysaccharide of interest in an extract of said cell, or the presence of a particularly glycosyltransferase activity in an extract of said cell; (ii) creating a genetic library of either cDNA or genomic DNA from the genetic material of said isolated cell; (iii) transforming host cells with said genetic library; and (iv) screening said transformed host cells for a host cell containing said post-translational characteristic, thereby obtaining a cell containing said gene, is disclosed. The method can be used to obtain genes encoding glycosyltransferases.
一种分离基因的方法,包括:(i)分离具有感兴趣的翻译后特征的细胞,该翻译后特征是所述细胞表面存在感兴趣的膜结合寡糖或多糖、感兴趣的可溶性寡糖或多糖提取物或者感兴趣的特定糖基转移酶活性,(ii)从所述分离的细胞的遗传物质中创建cDNA或基因组DNA的基因库,(iii)将所述基因库转化为宿主细胞,(iv)筛选所述转化的宿主细胞以获取包含所述翻译后特征的宿主细胞,从而获得包含所述基因的细胞。该方法可用于获得编码糖基转移酶的基因。