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US 2022/0177927 A1Published application
Method for producing transformant
AssigneeToyota Jidosha Kabushiki Kaisha
InventorToru Onishi
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Abstract
The present disclosure concerns evaluation as to whether or not a nucleic acid fragment having a target gene had been accurately integrated into the host genome. A group of nucleic acid fragments comprising a nucleic acid fragment having a target gene is introduced into host cells, and host cells in which the target gene had been cleaved from the genome DNA by the action of a site-specific recombinase are selected.
Claims4
A method for producing a transformant comprising steps of:
- introducing a group of nucleic acid fragments comprising a nucleic acid fragment having a target gene, a nucleic acid fragment having either one of the pair of homologous recombination sequences corresponding to a particular region of genome DNA, and a nucleic acid fragment having the other of the pair of homologous recombination sequences into a host cell;
- and
- selecting a host cell in which the target gene flanked by the pair of recognition sequences recognized by a site-specific recombinase had been cleaved from genome DNA,
- wherein nucleic acid fragments constituting the group of nucleic acid fragments can be linked to each other via a homologous recombination sequence provided at the end of each nucleic acid fragment, and
- wherein at least one of the pair of recognition sequences is provided in a nucleic acid fragment other than the nucleic acid fragment having a target gene among the group of nucleic acid fragments.
2 more elements
Dependencies are inferred from the claim text.