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US 9,315,830 B2Granted
Method of producing a recombinant microorganism
AssigneeLanzatech New Zealand Limited
InventorsDavid Jeffrey Fraser Walker, Michael Koepke
Life of this patent
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Abstract
The invention provides a genetic tool to insert, replace, delete, or otherwise manipulate a nucleic acid sequence in a microorganism to produce a recombinant microorganism. Notably, the invention makes use of homologous recombination, a type of genetic recombination in which nucleotide sequences are exchanged between two similar or identical molecules of DNA. Since the invention involves three homologous recombination events, it is referred to as a “triple cross” method.
Claims18
A method of producing a recombinant microorganism, comprising:
- (a) providing a microorganism comprising a genetic element comprising target nucleic acid T1, a target nucleic acid T2, and a target nucleic acid T3,
- (b) providing a DNA construct comprising a left homology arm LHA1 homologous to T1, a right homology arm RHA1 homologous to T2, and a right homology arm RHA2 homologous to T3, wherein RHA2 is located between LHA1 and RHA1,
- (c) allowing the genetic element of (a) to undergo homologous recombination with the DNA construct of (b), whereby T1 aligns with LHA1 and T2 aligns with RHA1 to insert the portion of the DNA construct between LHA1 and RHA1, including RHA2, into the genetic element between T1 and T2, and
- (d) allowing the genetic element of (c) to undergo self-homologous recombination, whereby T3 aligns with RHA2 to remove the portion of the genetic element between T3 and RHA2.
1 more element
Dependencies are inferred from the claim text.