ΦC31 Integrase
Product Introduction
ΦC31 integrase is derived from the integrase of Streptomyces phage phiC31. It is a large serine recombinase belonging to the serine recombinase family. This enzyme plays a key role in the integration of the phage genome and is encoded by phage phiC31. It efficiently catalyzes site-specific recombination between two specific sequences: the attP site in the phage genome and the attB site in the host bacterial chromosome.
The most distinctive feature of ΦC31 integrase lies in its unique recombination mechanism—by recognizing and recombining the two heterotypic sites attP and attB, it generates the product sites attL and attR, which are no longer substrates for this integrase. Thus, the integration is unidirectional and essentially irreversible. This characteristic distinguishes it from classical recombinases such as Cre and Flp (which recognize homotypic sites), making it an ideal tool for site-specific genomic integration.
ΦC31 integrase is composed of a protein of approximately 68 kDa (coding sequence length of about 1.8–2.4 kb). Studies have shown that this enzyme functions in both prokaryotic and eukaryotic systems, can integrate plasmids of any size (as single copies) without the need for cofactors, and the transgene remains stably expressed after integration.
Product Specifications
Parameter | Specification |
Source | Recombinant expression in E. coli |
Molecular Weight | ~62 kDa |
Concentration | 2.6 mg/ml |
Activity | Mediates site-specific recombination between attP and attB sites, enabling unidirectional DNA integration |
Purity | ≥95% (SDS-PAGE) |
Endotoxin | <1 EU/μg |
Storage Buffer | 50 mM Tris-HCl, 500 mM NaCl, 50% Glycerol, pH 8.0 |
10× Reaction Buffer | 200 mM Tris-HCl, 1 M NaCl, 1 mM EDTA, pH 7.5 |
Storage Conditions | Long-term storage at -80°C; short-term storage at -20°C |
Product Sizes
Specifications | Catalog Number | Concentration | Volume |
50 μL | GR300401 | 2.6 mg/mL | 50 μL |
250 μL | GR300402 | 2.6 mg/mL | 250 μL |
500 μL | GR300403 | 2.6 mg/mL | 500 μL |
Application Scenarios
Site-Specific Transgene Integration: By inserting the attB site into a donor plasmid and the attP site into the host genome, single-copy site-specific integration is achieved. It can integrate DNA fragments of any size, making it an ideal solution for establishing stable cell lines.
Non-Viral Gene Delivery and Gene Therapy: ΦC31 integrase enables efficient gene delivery without viral vectors, avoiding the immunogenicity and packaging limitations of viral vectors. Preclinical studies in systems such as liver, lung, neural stem cells, muscle stem cells, umbilical cord epithelial cells, and induced pluripotent stem cells have made progress, offering new strategies for gene therapy.
Minicircle DNA Preparation: In minicircle DNA technology, ΦC31 integrase recognizes attP/attB sites to recombine the parent plasmid, producing a minicircle plasmid containing only the eukaryotic expression elements and the gene of interest, used for efficient and safe transgenic vector construction.
Cell Engineering and Stable Cell Line Development: ΦC31 integrase is used in cell engineering platforms such as Jump-In™, enabling the generation of stable cell lines within two weeks, significantly shortening the development cycle compared to traditional methods. Combined with R4 integrase, it allows isogenic protein expression from defined genomic loci, suitable for comparative analysis of gene families, splice isoforms, or orthologs.
Gene Editing in Model Organisms: ΦC31 integrase has been widely used for site-specific integration and gene knockout in model organisms such as Drosophila. attP-containing "docking lines" enable efficient integration of attB-tagged donor DNA into specific genomic sites.
Plant Synthetic Biology: ΦC31 integrase has been used to construct synthetic biology circuits such as memory switches in plants, providing tools for modifying plant traits, introducing disease resistance, and enhancing nutritional content.
References
1. Groth, A. C., et al. (2000). A phage integrase directs efficient site-specific integration in human cells. Proceedings of the National Academy of Sciences, 97(11), 5995–6000.
2. Thorpe, H. M., & Smith, M. C. (1998). In vitro site-specific integration of bacteriophage DNA catalyzed by a recombinase of the resolvase/invertase family. Proceedings of the National Academy of Sciences, 95(10), 5505–5510.
3. Thyagarajan, B., et al. (2001). Site-specific genomic integration in mammalian cells mediated by phage phiC31 integrase. Molecular and Cellular Biology, 21(12), 3926–3934.
4. Bernabe-Orts, J. M., Quijano-Rubio, A., Vazquez-Vilar, M., et al. (2020). A memory switch for plant synthetic biology based on the phage φC31 integration system. Nucleic Acids Research, 48(6), 3379–3394.
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