pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins.
Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal.
The vectors (Figure 1) offer the following features:
Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
Cytomegalovirus (CMV) promoter for high-level constitutive expression
C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
C-terminal c-myc epitope for detection with an Anti-myc Antibody
Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)
The pSecTag2 vectors carry the Zeocin resistance gene for cost-effective selection in mammalian cells. Zeocin selection can also be used in E. coli.
The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.
引用及參考文獻
Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10.
Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM, Sekiguchi K, J Biol Chem (2004) 279:10946-10954
Product usage: The 5 cDNA was inserted into the NheI/PmeI sites of pcDNA3.1. For site-directed mutagenesis of the LG3 module, the cDNA fragment encoding LG3 was excised from pcDNA-5LG4–5 with AscI and PmeI and recloned into pSecTag2A at the AscI/PmeI sites. Recombinant protein was produced usi
Prohaptoglobin is proteolytically cleaved in the endoplasmic reticulum by the complement C1r-like protein.Wicher KB, Fries E, Proc Natl Acad Sci U S A (2004) 101:14390-14395
Product usage: HEK293 and COS-1 cells were transiently transfected with C1r-LP cDNA cloned into pcDNA6-myc-his or with proHp cDNA cloned into pSecTag. After 24 h, the cells were rinsed twIce with PBS and grown for 48 h in a serum-free medium. The cell media were then collected.
