US 12,351,805 B2
Method of making recombinant silk and silk-amyloid hybrid proteins using bacteria
Neel Satish Joshi, Somerville, MA (US); Peter Quoc Nguyen, Garden Grove, CA (US); Noemie-Manuelle Dorval Courchesne, Montreal (CA); and Zahra Abdali, Montreal (CA)
Assigned to Northeastern University, Boston, MA (US); and The Royal Institution for the Advancement of Learning/McGill University, Montréal (CA)
Filed by Northeastern University, Boston, MA (US); and The Royal Institution for the Advancement of Learning/McGill University, Montréal (CA)
Filed on May 14, 2021, as Appl. No. 17/320,866.
Application 17/320,866 is a continuation in part of application No. 15/776,998, granted, now 11,365,225, previously published as PCT/US2016/062820, filed on Nov. 18, 2016.
Claims priority of provisional application 62/257,405, filed on Nov. 19, 2015.
Prior Publication US 2021/0380992 A1, Dec. 9, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 4/04 (2006.01); C07K 1/22 (2006.01); C07K 1/30 (2006.01); C07K 1/34 (2006.01); C07K 14/245 (2006.01); C08J 3/075 (2006.01); C08J 5/18 (2006.01); C12N 1/20 (2006.01); C12N 15/62 (2006.01); C12N 15/70 (2006.01); C12R 1/19 (2006.01)
CPC C12N 15/70 (2013.01) [C07K 1/22 (2013.01); C07K 1/30 (2013.01); C07K 1/34 (2013.01); C07K 14/245 (2013.01); C12N 1/20 (2013.01); C07K 2319/02 (2013.01); C07K 2319/21 (2013.01); C12R 2001/19 (2021.05)] 20 Claims
 
1. A method of producing a genetically modified Escherichia coli (E. coli) bacterium, comprising
genetically altering the bacterium to include a nucleic acid sequence encoding a recombinant bacterial collagen protein comprising a Sec domain for periplasmic localization and an N22 domain for directing the recombinant bacterial collagen protein to the outer membrane for secretion;
wherein the bacterial collagen protein comprises a trypsin-sensitive globular variable domain comprising the amino acid sequence set forth in SEQ ID NO: 2075, and at least one collagen domain comprising the amino acid sequence set forth in SEQ ID NO: 2076;
wherein the nucleic acid is under operation of a promoter to express the recombinant bacterial collagen protein.