Schäfer K, Braun T
Dept. of Cell and Molecular Biology, University of Braunschweig, FRG.
Biochem Biophys Res Commun. 1995 Feb 15;207(2):708-14. doi: 10.1006/bbrc.1995.1245.
The FLAG peptide has been widely used as a multi-purpose tag for the identification and detection of recombinant FLAG fusion proteins. The practicability of this approach depends on specific detection of FLAG fusion proteins with no or very little cross-reactivity to cellular proteins. We have isolated a rat cDNA clone coding for a new splicing isoform of Mg2+ dependent protein phosphatase beta (MPP beta) by screening a rat brain expression library with monoclonal antibody Anti-FLAG M2. MPP beta reacts strongly both as a MPP beta-beta-galactosidase- and as a glutathione S-transferase fusion protein with anti-FLAG M2 antibodies. Sequence analysis of MPP beta revealed a sequence motif with five out of eight amino acid residues identical to the FLAG peptide hitherto believed to be mono-specific.
FLAG肽已被广泛用作多功能标签,用于鉴定和检测重组FLAG融合蛋白。这种方法的实用性取决于对FLAG融合蛋白的特异性检测,且与细胞蛋白的交叉反应极少或没有。我们通过用单克隆抗体抗FLAG M2筛选大鼠脑表达文库,分离出了一个编码Mg2+依赖性蛋白磷酸酶β(MPPβ)新剪接异构体的大鼠cDNA克隆。MPPβ作为MPPβ-β-半乳糖苷酶融合蛋白和谷胱甘肽S-转移酶融合蛋白时,均与抗FLAG M2抗体发生强烈反应。MPPβ的序列分析揭示了一个序列基序,其中八个氨基酸残基中有五个与迄今认为具有单特异性的FLAG肽相同。