A novel gene amplification system in yeast based on double rolling-circle replication.

Double rolling circle replication (DRCR) is recombinogenic

Homologous recombination plays a critical role in maintaining genetic diversity as well as genome stability. Interesting examples implying hyper-recombination are found in nature. In chloroplast DNA (cpDNA) and the herpes simplex virus 1 (HSV-1) genome, DNA sequences flanked by inverted repeats undergo inversion very frequently, suggesting hyper-recombinational events. However, mechanisms respo...

Gene amplification system based on double rolling-circle replication as a model for oncogene-type amplification

Gene amplification contributes to a variety of biological phenomena, including malignant progression and drug resistance. However, details of the molecular mechanisms remain to be determined. Here, we have developed a gene amplification system in yeast and mammalian cells that is based on double rolling-circle replication (DRCR). Cre-lox system is used to efficiently induce DRCR utilizing a rec...

Double Rolling Circle Replication (DRCR): Involvement in Gene Amplification and Genome Replication including HSV

Direct retransformation of yeast with plasmid DNA isolated from single yeast colonies using rolling circle amplification.

We have efficiently amplified plasmid DNA from single yeast colonies using rolling circle amplification (RCA). The amplified DNA can be directly used for restriction digestion, DNA sequencing, or yeast transformation. The RCA-based high-fidelity amplification would be useful for plasmid manipulation in a variety of yeast-based systems, particularly for high-throughput analyses.

Rtt109 Prevents Hyper-Amplification of Ribosomal RNA Genes through Histone Modification in Budding Yeast

The genes encoding ribosomal RNA are the most abundant in the eukaryotic genome. They reside in tandem repetitive clusters, in some cases totaling hundreds of copies. Due to their repetitive structure, ribosomal RNA genes (rDNA) are easily lost by recombination events within the repeated cluster. We previously identified a unique gene amplification system driven by unequal sister-chromatid reco...