Cloning and Protein Expression
|There are many options available for protein expression from cloned DNA. These include cell-free extracts (in vitro expression systems), and bacterial, yeast, insect, and mammalian cell systems, each with its own advantages and drawbacks. Requirements for protein solubility, functionality, and yield are often the most important factors to consider when choosing an expression system, but the time and labor required for subcloning and optimizing expression can also have a significant impact. |
Use of recombinant proteins varies widely - from functional studies in vivo to large-scale protein production for structural studies and therapeutics. Using the best expression system for your protein and application is key to your success.
The following table highlights the characteristics of the commonly-used expression hosts.
Select the best expression host for your research
|Host organism||Typical applications||Advantages||Potential Challenges|
|Cell-free|| || || |
|Bacteria|| || || |
|Yeast|| || || |
|Insect|| || || |
|Mammalian|| || || |
Protein Expression with Gateway® technology
The Gateway® recombination cloning system is designed for highly efficient transfer of your DNA into any Gateway® expression vector from a single entry clone. The extremely wide variety of Gateway® cloning–compatible expression vectors available makes the Gateway® system of recombination cloning ideal for protein expression studies.
- One hour, room temperature cloning reactions with up to >99% efficiency deliver the clone you need
- Maintain orientation and reading frame without using restriction enzymes or ligation to make expression-ready clones
- Use the same clone from target identification to validation to help ensure consistent results throughout your experiment, without re-sequencing
- Easily shuttle insert DNA from one expression vector to another—get flexibility while simplifying your cloning workflow
- Learn more about Gateway® cloning
For T7-regulated E. coli expression systems, MagicMedia™ Expression Medium can increase protein yields up to 10-fold. The exclusive MagicMedia™ formulation enables E. coli to reach culture densities 3–10-fold higher than those achieved with traditional LB + IPTG media.
- Learn more about Magic™Media medium