Change search
ReferencesLink to record
Permanent link

Direct link
A structure-function analysis of P2 integras
Stockholm University, Faculty of Science, Department of Genetics, Microbiology and Toxicology.
(English)Manuscript (Other academic)
URN: urn:nbn:se:su:diva-25721OAI: diva2:200373
Part of urn:nbn:se:su:diva-8502Available from: 2009-02-12 Created: 2009-02-03 Last updated: 2010-05-04Bibliographically approved
In thesis
1. Site-specific recombination in P2-like coliphages
Open this publication in new window or tab >>Site-specific recombination in P2-like coliphages
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The scope of these studies has been to investigate the site-specific recombination systems of P2-like coliphages, both in an evolutionary perspective by a comparative analysis of related phages as well as in a functional perspective.

Surveys of P2-like phages in Escherichia coli isolated from nature reveal the existence of seven discrete immunity classes and three integration sites, one of them previously unknown. Phylogenetic analysis of the repressor proteins and other analyses show that homologous recombination plays a role in the appearance of new immunities. Other studies of P2-like prophages from sequenced genomes from public databases show that the P2-like phages grow in different γ-proteobacteria. Based on the type of immunity and site-specific recombination system they can be roughly subdivided in two distinct subgroups and some new host integration sites could be identified. Some of the host attachment sites have a high identity to the sequences in the human genome, making them interesting as potential tools for targeted gene insertions into unmodified human cells.

The functional studies have been focused on the identification of the determinants for site specificity, which is important for the use of the enzyme for targeted gene insertions into unmodified genomes. Two approaches have been used. In one, we have performed a structure-function analysis of P2 Int that has identified several presumptive residues involved in specific binding to the core sequence, all of them located in the same alpha-helix. This knowledge could be a base for an in vitro evolution of the integrase to enable it to accept new DNA targets with a high affinity. With respect to the excisionases from P2-like coliphages integrating in different sites, we found that they share some common features when they bind and bend to their DNA targets, but there are also significant differences, especially those related to the number of binding sites and the distribution of these and the IHF binding sites in the attP regions. In the other approach we have started to characterize the site-specific recombination system of another P2-like phage, ΦD145, that has a host target with a high identity to a site in the human genome. This looks promising since the human sequence can be used in vivo in E. coli with a rather high efficiency.

Place, publisher, year, edition, pages
Stockholm: Institutionen för genetik, mikrobiologi och toxikologi, 2009. 41 p.
National Category
Research subject
Molecular Genetics
urn:nbn:se:su:diva-8502 (URN)978-91-7155-817-6 (ISBN)
Public defence
2009-03-06, sal E306, Arrheniuslaboratorierna, Svante Arrhenius väg 14-18, Stockholm, 10:00 (English)
Available from: 2009-02-12 Created: 2009-02-03 Last updated: 2012-02-28Bibliographically approved

Open Access in DiVA

No full text

Search in DiVA

By author/editor
Sylwan, L.
By organisation
Department of Genetics, Microbiology and Toxicology

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 41 hits
ReferencesLink to record
Permanent link

Direct link