	US 11,702,665 B2
	Paenibacillus-based endospore display platform, products and methods
Damian Curtis, Davis, CA (US); Benjamin L. Golomb, Sacramento, CA (US); Dilara Ally, Natick, MA (US); Florencia A. Ficarra, Davis, CA (US); Rauf Salamzade, Madison, WI (US); and Bjorn A. Traag, Walnut Creek, CA (US)
Assigned to Ginkgo Bioworks, Inc., Boston, MA (US)
Filed by Ginkgo Bioworks, Inc., Boston, MA (US)
Filed on Mar. 30, 2021, as Appl. No. 17/217,952.
Application 17/217,952 is a division of application No. 16/191,964, filed on Nov. 15, 2018, granted, now 10,988,769, issued on Apr. 27, 2021.
Claims priority of provisional application 62/587,371, filed on Nov. 16, 2017.
Prior Publication US 2021/0238610 A1, Aug. 5, 2021
Int. Cl. C12N 15/74 (2006.01); C12N 9/04 (2006.01); C07K 14/325 (2006.01); C12N 9/24 (2006.01); C12N 9/42 (2006.01); C07K 14/195 (2006.01); C12N 11/16 (2006.01); C12N 15/62 (2006.01); C12N 1/20 (2006.01); C12N 15/10 (2006.01); A01N 63/50 (2020.01); A01N 63/10 (2020.01); A01N 63/00 (2020.01)

CPC C12N 15/74 (2013.01) [A01N 63/00 (2013.01); A01N 63/10 (2020.01); A01N 63/50 (2020.01); C07K 14/195 (2013.01); C07K 14/325 (2013.01); C12N 1/20 (2013.01); C12N 9/0006 (2013.01); C12N 9/24 (2013.01); C12N 9/2442 (2013.01); C12N 11/16 (2013.01); C12N 15/1037 (2013.01); C12N 15/625 (2013.01); C12Y 101/01004 (2013.01); C12Y 302/01014 (2013.01); C07K 2319/035 (2013.01)]

7 Claims

1. A method for identifying an N-terminal signal sequence that is capable of targeting a protein to a spore surface of an endospore-forming bacterium, comprising:

screening a genome of the endospore-forming bacterium for at least one open reading frame which encodes a protein having multiple collagen-like triplet repeats having the sequence “GXX” wherein “X” represents “any amino acid”;

determining that at least one of the proteins identified in the screening step localizes to the spore surface of the endospore-forming bacterium by microscopy or experimentally; and

identifying a putative N-terminal signal sequence from at least one protein identified in the determining step as localizing to the spore surface and expressing in the endospore-forming bacterium a fusion protein comprising the putative N-terminal signal sequence and a reporter gene.