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The Penn State Protein Ladder system for inexpensive protein molecular weight markers
Ryan T Santilli1,2, John E Williamson1, Yoshitaka Shibata1,2
1Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, 16802, USA.
Scientific Reports
|August 19, 2021
Summary
Researchers developed the Penn State Protein Ladder system for affordable protein molecular weight markers. This system uses engineered E. coli plasmids to easily produce and detect various protein sizes for research and teaching applications.
Area of Science:
- Molecular Biology
- Biochemistry
- Protein Expression
Background:
- Standard protein molecular weight markers can be expensive.
- There is a need for cost-effective and easily producible protein ladder systems.
Purpose of the Study:
- To develop an inexpensive and user-friendly system for producing protein molecular weight markers.
- To create expression plasmids for a range of protein sizes in E. coli.
Main Methods:
- Engineered E. coli expression plasmids for proteins of 10-250 kD.
- Utilized histidine tags for easy purification and engineered immunoglobulin binding domains for Western blot detection.
- Developed polycistronic vectors for coexpression of multiple proteins.
Main Results:
- Achieved high protein expression levels (10-50 mg/L) suitable for SDS-PAGE and Western blotting.
- Demonstrated cost-effective production (less than a penny per lane).
- Showcased efficient production of numerous ladder lanes from small culture volumes.
Conclusions:
- The Penn State Protein Ladder system offers an economical and efficient solution for protein molecular weight markers.
- The system serves as a valuable tool for both research and educational laboratories.
- Facilitates recombinant protein expression, purification, and detection applications.