Screening for proteins interacting with MCM7 in human lung cancer library using yeast two hybrid system
Background and objective 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 containi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | zho |
| Published: |
Chinese Anti-Cancer Association; Chinese Antituberculosis Association
2008-08-01
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| Series: | Chinese Journal of Lung Cancer |
| Subjects: | |
| Online Access: | http://www.lungca.org/index.php?journal=01&page=article&op=view&path%5B%5D=10.3779%2Fj.issn.1009-3419.2008.04.012&path%5B%5D=116 |
| Summary: | Background and objective 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. Conclusion The investigation provides functional clues for further exploration of MCM7 gene. |
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| ISSN: | 1009-3419 1999-6187 |