BMB Reports
- Volume 32 Issue 5
- /
- Pages.497-501
- /
- 1999
- /
- 1976-670X(eISSN)
Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli
- Cho, Eun-Wie (Peptide Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology) ;
- Park, Jung-Hyun (Peptide Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology) ;
- Na, Shin-Young (Peptide Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology) ;
- Kim, Kil-Lyong (Peptide Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology)
- Received : 1999.04.24
- Accepted : 1999.05.15
- Published : 1999.09.30
Abstract
Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-