Related Experiment Video
Updated: Mar 6, 2026
Author Spotlight: Expression and Purification of Human Solute Carrier Transporters Using Codon-Optimized Genes
Published on: September 29, 2023
Optimized protocol for soluble prokaryotic expression, purification and structural analysis of human placenta
Mahboobeh Nazari1, Amir-Hassan Zarnani2, Roya Ghods3
1Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Department of Tissue Engineering, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Producing soluble Placenta specific -1 (PLAC1) protein was challenging due to disulfide bridges. This study optimized recombinant PLAC1 production using an expression matrix, yielding soluble truncated PLAC1 and improving full-length protein recovery.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Expression
Background:
- Placenta specific -1 (PLAC1) is a membrane-associated protein crucial for placental development.
- PLAC1 expression is detected in numerous cancer cell lines, suggesting potential roles in oncology.
- Disulfide bridges in PLAC1 hinder soluble recombinant protein production and structural studies.
Purpose of the Study:
- To optimize the production of soluble full-length and truncated human PLAC1.
- To establish an efficient system for generating recombinant PLAC1 for further research.
Main Methods:
- Utilized an expression matrix involving vectors, E. coli hosts, and solubilization conditions.
- Characterized recombinant proteins via Western blotting (WB) and enzyme-linked immunosorbent assay (ELISA).
- Investigated protein structure using circular dichroism (CD) spectroscopy.
Main Results:
- Optimized conditions yielded soluble truncated PLAC1 using Origami™ and pCold vector without additional solubilization.
- Full-length PLAC1 predominantly formed inclusion bodies, with higher yields in Origami™ and Rosetta2 hosts.
- A buffer with 2M Urea at pH 12 proved effective for solubilizing full-length PLAC1.
- Recombinant PLAC1 proteins showed high reactivity in WB and ELISA assays.
Conclusions:
- Developed an efficient and cost-effective expression system for soluble PLAC1.
- The optimized system facilitates further investigation into PLAC1's physiological roles and potential applications.
