Development of a new epitope tag recognized by a monoclonal antibody to Rickettsia typhi
Development of a new epitope tag recognized by a monoclonal antibody to Rickettsia typhi
复制标题
DOI：
10.2144/01313st08
复制
发表时间：
2001-09-01
期刊：
影响因子：
2.7
通讯作者：
Hahn, MJ
中科院分区：
文献类型：
--
作者：
Lee, JR;Chang, YY;Hahn, MJ
鼠抗伤寒立克次体晶体表层蛋白的单抗SRT10识别的表位被定位为10个氨基酸残基(SRTag TFIGAIATDT)。将覆盖SRT10识别表位的寡核苷酸序列与多个克隆位点一起插入哺乳动物表达载体。将SRTag与NCC27/CLIC1基因编码区融合，在哺乳动物细胞中表达，通过免疫印迹、免疫细胞化学和免疫沉淀等方法检测到标记蛋白。除SRT-NCC27/CLIC1外，SRT10还能通过免疫细胞化学检测到N端标记的MEF2D和C端标记的CD4。我们认为SRT10对SRTag的这种特异性识别普遍适用于需要表达和检测融合蛋白的细胞和分子生物学研究。
The epitope recognized by a mouse monoclonal antibody (MAb) to the crystalline surface layer protein of Rickettsia typhi, SRT10, was mapped to 10 amino acid residues (SRTag TFIGAIATDT). The oligonucleotide sequence covering the epitope recognized by SRT10 was inserted into a mammalian expression vector together with multiple cloning sites. When the SRTag was fused in frame to the coding region of the NCC27/CLIC1 gene and expressed in mammalian cells, the MAb SRT10 could detect the tagged protein by immunoblotting, immunocytochemistry, and immunoprecipitation. In addition to the SRT-NCC27/CLIC1, SRT10 could detect N-terminal-tagged MEF2D and C-terminal-tagged CD4 by immunocytochemistry. We suggest that this specific recognition of the SRTag by SRT10 is generally applicable to cellular and molecular biology research that requires the expression and detection of fusion proteins.