| 主权项 |
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. |
