ColE1
This article needs additional citations for verification. (January 2017) |
ColE1 is a plasmid found in bacteria. Its name derives from the fact that it carries a gene for colicin E1 (the cea gene). It also codes for immunity from this product with the imm gene. In addition, the plasmid has a series of mobility (mob) genes. Its size and the presence of a single EcoRI recognition site caused it to be considered as a vector candidate.[1]
Replication
ColE1 replication begins at the origin. 555bp upstream from this point, RNA polymerase initiates transcription of RNAII which acts as a pre-primer and begins the synthesis of the leader strand. The transcript folds into a secondary structure which stabilises the interaction between the nascent RNA and the origin's DNA. This hybrid is attacked by RNase H, which cleaves the RNA strand, exposing a 3' hydroxyl group. This allows the extension of the leading strand by DNA polymerase I. Lagging strand synthesis is later initiated by a primase encoded by the host cell. Replication is carried out entirely by host proteins (RNA polymerase, DNA polymerase I and RNase H) so that inhibition of translation will stop the growth of the cells, but not the replication of ColE1. Since the translation of Rop protein will also be inhibited, a higher than normal copy number will result in these cells.
Copy number control
RNAI is a counter-transcript to a section of RNAII and so binds to its 5' end. This alters the folding of RNAII so that the DNA-RNA hybrid is not stabilized and cleavage does not occur. This ensures that at high copy numbers, replication is slowed down due to increased RNAI concentration. Rop is a secondary replication repressor, it stabilizes the RNAI-RNAII hybrid. Rop may be especially important at preventing runaway replication at slow growth rates.

ColE1 has a copy number of 25-30 according to its original publication.[1]
Related plasmids and compatibility
The copy-number control mechanism affects all plasmids with the same type of mechanism as long as their RNAIs are similar enough. As a result, two different plasmids with similar RNAIs cannot stably co-exist; this is called incompatibility. Natural plasmids sharing the ColE1 mechanism form at least four incompatibility groups (ColE1, p15A, CloDF13, RSF1030). Plasmids from different groups are compatible with each other because they do not overly interfere with each other. Directed evolution has been used to generate even more incompatibility groups.[2]
References
- ^ a b Hershfield, Vickers; Boyer, Herbert W.; Yanofsky, Charles; Lovett, Michael A.; Helinski, Donald R. (September 1974). "Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNA". Proceedings of the National Academy of Sciences. 71 (9): 3455–3459. Bibcode:1974PNAS...71.3455H. doi:10.1073/pnas.71.9.3455. PMC 433792. PMID 4610576.
- ^ Chaillou, S; Stamou, PE; Torres, LL; Riesco, AB; Hazelton, W; Pinheiro, VB (9 September 2022). "Directed evolution of colE1 plasmid replication compatibility: a fast tractable tunable model for investigating biological orthogonality". Nucleic Acids Research. 50 (16): 9568–9579. doi:10.1093/nar/gkac682. PMC 9458437. PMID 36018798.
Content Disclaimer
Informasi ini disarikan dari Wikipedia dan disajikan kembali untuk tujuan edukasi. Konten tersedia di bawah lisensi CC BY-SA 3.0. Kami tidak bertanggung jawab atas ketidakakuratan data yang bersumber dari kontribusi publik tersebut.
- The information displayed on this website is sourced in part or in whole from Wikipedia and has been adapted for the purpose of restating it. We strive to provide accurate and relevant information, however:
- There is no guarantee of absolute accuracy. Wikipedia is an open, collaborative project that can be edited by anyone, so information is subject to change.
- It is not intended to constitute professional advice. The content displayed is for informational and educational purposes only. For important decisions (e.g., medical, legal, or financial), please consult a professional.
- Content copyright. Wikipedia is licensed under the Creative Commons Attribution-ShareAlike License (CC BY-SA). This means that content may be reused with appropriate attribution and shared under a similar license.
- Responsible use. Any risk arising from the use of information from this website is entirely the responsibility of the user.