Improved Cloning Strategy for High Throughput Expression of Eukaryotic Proteins
Author Information
Author(s): Chen Yunjia, Qiu Shihong, Luan Chi-Hao, Luo Ming
Primary Institution: University of Alabama at Birmingham
Hypothesis
Can a new domain/domain boundary prediction method combined with an improved cloning strategy enhance the expression of higher eukaryotic proteins?
Conclusion
The DDBP method and improved cloning strategy significantly increased the success rate of obtaining purified proteins.
Supporting Evidence
- The DDBP method was trained using available experimental data.
- More than 97% of targeted genes were obtained as entry clones.
- 50 out of 57 proteins were expressed as soluble after applying the DDBP method.
Takeaway
The researchers created a new way to find the best parts of proteins to clone, which helped them get more proteins that can be used for experiments.
Methodology
The study used a high throughput cloning pipeline that included bioinformatics tools for domain prediction and an automated robotic platform for cloning and expression.
Limitations
The study does not clarify whether the improvement in protein expression is solely due to correct domain prediction.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website