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US 10,415,043 B2Granted
Vitamin prototrophy as a selectable marker
AssigneeLanzatech New Zealand Limited
InventorsMichael Koepke, Bakir Al-Sinawi
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Abstract
One or more genes in a biosynthesis pathway for a vitamin or other essential nutrient which is needed for the survival of a microorganism can be used as an effective selective marker to identify cells transformed with an exogenous nucleic acid. The microorganism does not naturally contain or express the one or more gene. This permits genetic manipulations to be performed. It permits lower cost fermentations to be performed. It permits production of the essential nutrient for subsequent commodity use.
Claims8
A method of using vitamin prototrophy as a selectable marker comprising:
- (a) transforming a microorganism auxotrophic for a vitamin with both a vitamin biosynthesis gene and a gene conferring a desired property, wherein the microorganism is not transformed with an antibiotic resistance gene and
- (b) culturing the microorganism in the absence of the vitamin,
- wherein growth of the microorganism in the absence of the vitamin indicates successful transformation of the microorganism with both the vitamin biosynthesis gene and the gene conferring a desired property,
- wherein the vitamin is thiamine, the vitamin biosynthesis gene encodes thiamine biosynthesis protein (ThiC) (EC 4.1.99.17) derived from Clostridium ragsdalei, and the microorganism is Clostridium autoethanogenum.
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Dependencies are inferred from the claim text.