Foreman P K, Davis R W
Department of Biochemistry, Beckman Center, Stanford University School of Medicine, CA 94305.
Gene. 1994 Jun 24;144(1):63-8. doi: 10.1016/0378-1119(94)90204-6.
A series of cloning vectors, designated YCpIF, was constructed to facilitate the conditional synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae. These vectors contain a translation start codon upstream from a multiple cloning site (MCS) in each of the three reading frames. Protein synthesis is under the control of the GAL1 promoter, which drives transcription when cells are grown on galactose-containing medium, but not when they are grown on glucose-containing medium. Different versions of the vectors contain four different commonly used selectable markers. In addition, YCpIF15, YCpIF16 and YCpIF17 contain a sequence encoding an epitope from influenza virus hemagglutinin upstream from the MCS. These vectors facilitate the addition of this epitope tag to the N terminus of any protein. The epitope is recognized by a commercially available monoclonal antibody.
构建了一系列名为YCpIF的克隆载体，以促进酿酒酵母中表位标记、截短和嵌合蛋白的条件合成。这些载体在三个阅读框中的每一个的多克隆位点（MCS）上游都有一个翻译起始密码子。蛋白质合成受GAL1启动子控制，当细胞在含半乳糖的培养基上生长时，该启动子驱动转录，但在含葡萄糖的培养基上生长时则不驱动转录。载体的不同版本包含四种不同的常用选择标记。此外，YCpIF15、YCpIF16和YCpIF17在MCS上游包含一个编码来自流感病毒血凝素表位的序列。这些载体便于将该表位标签添加到任何蛋白质的N端。该表位可被市售单克隆抗体识别。
