User manual for Takara's pCold™ TF DNA v201910Da, a cold shock expression vector for soluble recombinant protein expression in E. coli.
This document provides detailed instructions for using the pCold™ TF DNA v201910Da vector to express recombinant proteins in E. coli. It includes information on vector structure, storage, usage methods, cloning procedures, and troubleshooting tips. The vector features a cold shock promoter (cspA), a Trigger Factor (TF) solubility tag, and cleavage sites for removing the tag. It is designed to improve protein solubility and yield compared to traditional expression systems.
The manual covers experimental examples, Q&A for common issues, and protocols for cloning and expression. It also discusses compatible E. coli hosts, optimization of expression conditions, and alternatives for difficult-to-express proteins. This manual is written in Chinese.
| Vector Name | pCold™ TF DNA |
|---|---|
| Promoter | cspA (cold shock protein A) |
| Solubility Tag | Trigger Factor (TF) |
| Molecular Weight of TF | 48 kDa |
| Cleavage Sites | HRV 3C Protease, Thrombin, Factor Xa |
| Host Compatibility | Most E. coli strains |
| Vector Size | 25 μg |
BL21, Rosetta, and Origami strains have been used. BL21 is the most commonly used host.
No, it is not recommended. Transformants should be stored as glycerol stocks at -80℃.
A 125 kDa human protein has been successfully expressed with good solubility.