pBeloBAC11 is a widely recognized bacterial artificial chromosome (BAC) vector that has played a fundamental role in genomic mapping, functional gene analysis, and large-insert cloning. Its robust design, high-fidelity replication in E. coli, and stable maintenance of large DNA fragments make it a staple in molecular biology laboratories.
What is pBeloBAC11?
pBeloBAC11 is derived from the F-plasmid origin of replication, enabling the stable maintenance of large DNA inserts (up to 300 kb) in a low-copy state within Escherichia coli host cells. It was developed as part of the broader effort to map and sequence complex genomes.
For a foundational reference, the vector design was described in detail by Shizuya et al., in their seminal paper (Proc Natl Acad Sci USA, 1992), laying the groundwork for BAC-based library construction.
Key Features of pBeloBAC11
- F-factor origin (oriS + repE): Ensures single-copy replication for stable inheritance.
- lacZα multiple cloning site (MCS): Supports blue-white screening for insert detection.
- Chloramphenicol resistance (Cm^R): Selection marker effective in both research labs and academic core facilities.
- T7 and SP6 promoters: Enable in vitro transcription of inserted sequences (source).
Explore more about vector maps and cloning strategies on NCBI’s GenBank.
Applications in Genomic Research
1. Human and Model Organism Genome Libraries
pBeloBAC11 has been instrumental in constructing BAC libraries for organisms such as:
- Homo sapiens.
- Mus musculus.
- Arabidopsis thaliana.
2. Functional Genomics and Transgenesis
The low-copy nature of BACs makes them ideal for producing transgenic models and conducting gene complementation studies.
pBeloBAC11-based constructs have been introduced into mammalian systems using electroporation and viral transduction methods described by the NIH Stem Cell Unit.
3. Structural Genomics and Chromatin Studies
BACs support the study of long-range regulatory elements and chromatin architecture through techniques like 3C, 4C, and Hi-C. These tools have revealed how gene enhancers and insulators interact over large genomic distances—insights made possible by stable BAC vectors like pBeloBAC11.
Technical Considerations
- Transformation efficiency is typically optimized using electrocompetent cells, such as DH10B or ElectroMAX™ strains.
- BAC DNA purification is best performed using kits recommended for low-copy plasmids.
- Stability of the insert should be verified using restriction enzyme mapping, pulse-field gel electrophoresis (PFGE), or next-generation sequencing.
The pBeloBAC11 vector remains a gold standard for stable, large-insert genomic cloning and functional genomic analysis. Its use in pioneering projects like the Human Genome Project and its ongoing application in chromatin research, genome engineering, and structural genomics highlight its value in modern biology.
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