1. A method for creating a self-replicating synthetic cell, said method comprising:
(i) assembling a synthetic bacterial, cyanobacterial, or microalgal donor genome as one or more fragments and introducing the donor genome as one or more fragments and a host vector into a yeast host cell, wherein the donor genome and the host vector are joined prior to or after introduction into the yeast host cell;
(ii) recovering the assembled donor genome from the yeast host cell;
(iii) performing step a) or b) or c) wherein a), b) and c) comprise
a) preparing the donor genome for transplantation into a bacterial, cyanobacterial, or microalgal recipient cell by methylating the donor genome;
b) preparing a bacterial or cyanobacterial or microalgal recipient cell by removing or inactivating a restriction endonuclease function present in the recipient cell that cuts the donor genome;
c) providing a bacterial, cyanobacterial, or microalgal recipient cell lacking a restriction endonuclease function that cuts the donor genome; and
(iv) introducing the recovered donor genome into the recipient cell,
thereby generating a self-replicating synthetic cell comprising the donor genome and controlled only by the donor genome, wherein the donor genome is sufficient to sustain viability and continuous self-replication of the recipient cell; and
wherein the synthetic cell supports gene expression from the donor genome and has a phenotype of the donor genome,
wherein the donor genome is an essentially intact genome that is at least a minimal genome, and is greater than about 300 kb in length.
2. The method of claim 1, wherein the donor genome and the host vector are introduced into the host cell simultaneously.
3. The method of claim 1, wherein the donor genome and host vector are joined prior to introduction into the yeast host cell by transforming the yeast host vector into a donor cell containing the donor genome.
4. The method of claim 3, wherein the host vector is a centromeric plasmid.
5. The method of claim 1, wherein the donor genome is modified within the yeast host cell.
6. The method of claim 1, further comprising degrading or removing the endogenous genome of the recipient cell.
7. The method of claim 1, wherein the recovered donor genome is methylated prior to introduction into the recipient cell.
8. The method of claim 1, wherein the recipient cell’s restriction endonuclease function is absent, removed or inactivated.
9. The method of claim 1, further comprising introducing a second donor genome into the host cell, wherein the second donor genome is different from the first donor genome, thereby producing a host cell containing two different donor genomes.
10. The method of claim 9, wherein introducing the second donor genome comprises mating the host cell containing the first donor genome with a second host cell containing the second donor genome.
11. The method of claim 1, wherein the synthetic cell exhibits a phenotype corresponding to the donor genome incorporating any modifications thereto.
12. The method of claim 1 wherein the synthetic donor genome is assembled in vitro prior to introducing the donor genome into the host cell.
13. The method of claim 5 wherein the modification of the donor genome is selected from the group consisting of a substitution, a deletion, an insertion, a rearrangement, and a recombination.
14. The method of claim 13 wherein the modification is selected from the group consisting of: an insertion, a deletion, and a substitution.
15. The method of claim 5 wherein the modification is a homologous recombination.
16. The method of claim 1 wherein the donor genome is a bacterial genome and the recipient cell is a bacterial cell.
17. The method of claim 1 wherein the donor genome is a cyanobacterial genome and the recipient cell is a cyanobacterial cell.
18. The method of claim 1 wherein the donor genome is a microalgal genome and the recipient cell is a microalgal cell.
19. The method of claim 1 wherein the yeast host cell is Saccharomyces cerevisiae or Saccharomyces pombe.
20. The method of claim 16, wherein the donor genome is a Mycoplasma genome and the recipient cell is a Mycoplasma cell.
21. The method of claim 19, wherein the recipient cell is a Mycoplasma capricolum.
22. The method of claim 20, wherein the recipient cell is a Mycoplasma capricolum.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A storage system comprising:
a first disk array system, in a primary site, coupled to a host computer, and having a plurality of first disk drive units in which data are stored and a first controller which controls to store data sent from said host computer to a primary volume related to a portion of said first disk drive units;
a second disk array system, in a remote site, coupled to said first disk array system, and having a plurality of second disk drive units in which data are stored and a second controller which receives data of said primary volume from said first disk array system by an asynchronous remote copy procedure and controls to store data received from said first disk array system to a secondary volume related to a portion of said second disk drive units; and
a third disk array system, coupled to said first disk array system, and having a plurality of third disk drive units in which data are stored and a third controller which receives data of said primary volume from said first disk array system by a synchronous copy procedure and controls to store data received from said first disk array system to a third volume related to a portion of said third disk drive units;
wherein said third disk array system comprises a first area and a second area, said first area storing information related to data of a plurality of write requests received from said first disk array system by said synchronous copy procedure during a first time period,
wherein said second area storing information related to data of a plurality of write requests received from said first disk array system by said synchronous copy procedure during a second time period being different from said first time period,
wherein said third disk array system stores information related to data of a plurality of write requests received from said first disk array system in said first area during said first time period and said second area during said second time period, and
wherein said information stored in said second area is used to decide data to be transferred from said third disk array system to said second disk array system in the event of a disaster that affects said primary site.