pBI-CMV5載體描述 pBI-CMV5 is a mammalian bidirectional expression vector designed to constitutively express a protein of interest and Metridia luciferase (MetLuc), a secreted reporter protein that can be easily detected in the medium surrounding the cells.
Metridia luciferase is a naturally secreted luciferase from the marine copepod Metridia longa. The human codon-optimized Metridia luciferase gene encodes a 219 amino acid (24 kDa) polypeptide that includes a 17 amino acid N-terminal signal peptide necessary for secretion (1). Metridia luciferase is expressed and secreted into the surrounding medium, where it is easily detected by the addition of a chemiluminescent substrate.
Protein expression is driven by one of two constitutively active, minimal human cytomegalovirus promoters: PminCMV1 (located upstream of the multiple cloning site [MCS]), drives the expression of the protein of interest, and PminCMV2 drives the expression of MetLuc. To allow propagation and selection in E. coli, the vector contains a ColE1 origin of replication and an ampicillin resistance gene (Ampr).
The pBI-CMV5 vector, available as part of the Bidirectional Secreted Luciferase System (Cat. No. 631757), is designed to constitutively express a protein of interest and Metridia luciferase. The gene of interest must contain an initiation codon and a stop codon.
pBI-CMV5 can be transfected into mammalian cells using any standard transfection method. Cells expressing the secreted Metridia luciferase can be detected by adding chemiluminescent substrate (provided) to a sample of the cell medium.
Propagation in E. coli
Recommended host strain: DH5α? and other general purpose strains.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
E. coli replication origin: ColE1
Copy number: low
Plasmid incompatibility group: pMB1/ColE1
References
1. Markova, S.V. et al. (2004) J. Bio. Chem. 279(5):3212-3217.
Note: The vector sequence was compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.
