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Updated: Jun 9, 2026
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
Published on: June 6, 2018
[Screening for proteins interacting with MCM7 in human lung cancer library using yeast two hybrid system.]
Yuchen Han1, Zhongyan Guan, Hongmei Duan
1Department of Pathology, College of Basic Medical Sciences and the First Affiliated Hospital, China Medical University, Shenyang, Liaoning 110001, China. hanyuchencn@yahoo.com.
Background:
MCM7 is a subunit of the MCM complex that plays a key role in DNA replication initiation. But little is known about its interaction proteins. In this study yeast two hybrid screening was used to identify the MCM7 interacting proteins.
Methods:
Yeast expression vector containing human full length MCM7-pGBKT7 plasmid was constructed, and with a library of cDNAs from human lung cancer-pACT2 plasmid was transformed into yeast strain AH109, and was electively grew in X-a-gal auxotrophy medium SD/-Trp-Leu-His-Ade, and the blue colonies were picked up, the plasmid of the yeast colonies was extracted , and transformed into E. Coli to extract DNA and performed sequence analysis.
Results:
Eleven proteins were identified which could specifically interact with MCM7 proteins, among these five were cytoskeleton proteins, six were enzymes, kinases and related receptors.
Conclusions:
The investigation provides functional clues for further exploration of MCM7 gene.
Insights
This study identified eleven proteins interacting with MCM7, a key DNA replication initiation factor. These findings offer new insights into MCM7
Area of Science:
- Molecular Biology
- Genetics
Context:
- The MCM complex, particularly the MCM7 subunit, is crucial for initiating DNA replication.
- Understanding MCM7's interaction partners is essential for elucidating its regulatory mechanisms.
- Limited information exists regarding proteins that interact with MCM7.
Purpose:
- To identify novel proteins that interact with the MCM7 subunit using a yeast two-hybrid screening approach.
- To expand the knowledge base of MCM7's functional network and its role in cellular processes.
Summary:
- A yeast two-hybrid screen was employed to identify MCM7 interacting proteins.
- Human full-length MCM7 was used with a human lung cancer cDNA library.
- Eleven interacting proteins were identified, including cytoskeleton proteins, enzymes, and kinases.
Impact:
- This research provides a foundation for further investigation into the functions of MCM7 and its associated proteins.
- The identified interaction partners may play roles in DNA replication, cell cycle regulation, or cancer development.
- These findings contribute to a deeper understanding of the MCM complex's role in genome stability.
