Development of a new epitope tag recognized by a monoclonal antibody to Rickettsia typhi

Citation
Jr. Lee et al., Development of a new epitope tag recognized by a monoclonal antibody to Rickettsia typhi, BIOTECHNIQU, 31(3), 2001, pp. 541-545
Citations number
7
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOTECHNIQUES
ISSN journal
0736-6205 → ACNP
Volume
31
Issue
3
Year of publication
2001
Pages
541 - 545
Database
ISI
SICI code
0736-6205(200109)31:3<541:DOANET>2.0.ZU;2-3
Abstract
The epitope recognized by a mouse monoclonal antibody (MAb) to the crystall ine surface layer protein of Rickettsia typhi, SRT10, was mapped to 10 amin o acid residues (SRTag TFIGAIATDT). The oligonucleotide sequence covering t he epitope recognized by SRT10 was inserted into a mammalian expression vec tor 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 ce lls, the MAb SRT10 could detect the tagged protein by immunoblotting, immun ocytochemistry, and immunoprecipitation. In addition to the SRT-NCC27/CLIC1 , SRT10 could detect N-terminal-tagged MEF2D and C-terminal-tagged CD4 by i mmunocytochemistry. 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.