Intravesicle Isothermal DNA Replication
1 Centre for Integrative Biology, University of Trento, Via delle Regole, 101 Mattarello, Italy
2 New England BioLabs, 240 County Road, Ipswich, Massachusetts 01938-2723, USA
3 School of Pharmacy, University of Colorado Denver, Aurora, Colorado 80045, USA
BMC Research Notes 2011, 4:128 doi:10.1186/1756-0500-4-128Published: 15 April 2011
Bacterial and viral DNA replication was previously reconstituted in vitro from component parts [1-4]. Significant advances in building minimal cell-like structures also have been made recently [5-7]. Combining the two approaches would further attempts to build a minimal cell-like structure capable of undergoing evolution by combining membrane encapsulation and genome replication. Towards this end, we attempted to use purified genomic replication protein components from thermophilic bacterial sources to copy strands of DNA isothermally within lipid vesicles.
Bacterial replication components (such as helicases and DNA polymerases) are compatible with methods for the generation of lipid vesicles. Encapsulation inside phospholipid vesicles does not inhibit the activity of bacterial DNA genome replication machinery. Further the described system is efficient at isothermally amplifying short segments of DNA within phospholipid vesicles.
Herein we show that bacterial isothermal DNA replication machinery is functional inside of phospholipid vesicles, suggesting that replicating cellular mimics can be built from purified bacterial components.