	US 12,391,627 B2
	Genetically modified nitrogen fixing bacteria and uses thereof
John Peters, Pullman, WA (US); Florence Mus, Pullman, WA (US); Jean-Michel Ane, Madison, WI (US); and Devanshi Khokhani, Madison, WI (US)
Assigned to WISCONSIN ALUMNI RESEARCH FOUNDATION, Madison, WI (US); and WASHINGTON STATE UNIVERSITY, Pullman, WA (US)
Filed by Wisconsin Alumni Research Foundation, Madison, WI (US); and Washington State University, Pullman, WA (US)
Filed on Sep. 18, 2020, as Appl. No. 17/024,746.
Claims priority of provisional application 63/056,050, filed on Jul. 24, 2020.
Claims priority of provisional application 63/018,551, filed on May 1, 2020.
Claims priority of provisional application 62/902,219, filed on Sep. 18, 2019.
Prior Publication US 2021/0107844 A1, Apr. 15, 2021
Int. Cl. C05F 11/08 (2006.01); C12N 1/20 (2006.01); C12N 15/74 (2006.01)

CPC C05F 11/08 (2013.01) [C12N 1/20 (2013.01); C12N 15/74 (2013.01)]

15 Claims

1. A genetically modified diazotrophic γ-proteobacterium exhibiting an increased ability to fix atmospheric nitrogen, comprising:

one or more insertions within the nifL gene of the wild type diazotrophic γ-proteobacterium, wherein the one or more insertions comprises a promoter sequence, and wherein the promoter sequence is placed and oriented to direct transcription of at least one gene of the rnf1 gene complex selected from the group consisting of rnfA1, rnfB1, rnfC1, rnfD1, rnfE1, rnfG1 and rnfH1 whereby the expression of the at least one gene of the rnf1 gene complex is upregulated relative to the wild type diazotrophic γ-proteobacterium;

wherein the one or more insertions disrupts the nifL gene.