Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a mammalian codon-optimized BirA biotin ligase.
Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a mammalian codon-optimized BirA biotin ligase.
复制标题
DOI：
10.1186/s13104-018-3500-9
复制
发表时间：
2018-06-14
影响因子：
1.8
通讯作者：
Strouboulis J
中科院分区：
文献类型：
--
作者：
Ioannou M;Papageorgiou DN;Ogryzko V;Strouboulis J
构建具有串联亲和标签的蛋白质的哺乳动物表达载体，所述串联亲和标签包括可生物素化的Avi标签和三重标记标签。我们通过瞬时转染法构建和测试了N-末端或C-末端标记蛋白的单个质粒的共表达，该载体携带由可生物素化的Avi标签和由烟草蚀刻病毒(TEV)蛋白酶切割位点分隔的三个FLAG标签组成的串联亲和标签，以及哺乳动物密码子优化的Bira生物素连接酶与绿色荧光蛋白的融合。我们还描述了用于任何选择的互补DNA的N-端或C-端AVI-TEV标记的平台载体。这些载体在哺乳动物细胞中核蛋白的代谢生物素化串联亲和标签的应用中提供了通用性和有效性。本文的在线版本(10.1186/s13104-0183500-9)包含补充材料，授权用户可以使用。
To construct mammalian expression vectors for the N- or C-terminal tagging of proteins with a tandem affinity tag comprised of the biotinylatable Avi tag and of a triple FLAG tag. We constructed and tested by transient transfections mammalian expression vectors for the co-expression from a single plasmid of N- or C-terminally tagged proteins bearing a tandem affinity tag comprised of the biotinylatable Avi tag and of a triple FLAG tag separated by a tobacco etch virus (TEV) protease cleavage site, together with a mammalian codon-optimized BirA biotin ligase fused to green fluorescent protein. We also describe platform vectors for the N- or C-terminal AVI-TEV-FLAG tagging of any complementary DNA of choice. These vectors offer versatility and efficiency in the application of metabolic biotinylation tandem affinity tagging of nuclear proteins in mammalian cells. The online version of this article (10.1186/s13104-018-3500-9) contains supplementary material, which is available to authorized users.