US 11,920,139 B2
Regulated expression of antigen and/or regulated attenuation to enhance vaccine immunogenicity and/or safety
Roy Curtiss, III, Gainesville, FL (US); Shifeng Wang, Gainesville, FL (US); Soo-Young Wanda, Gainesville, FL (US); and Wei Kong, Phoenix, AZ (US)
Assigned to The Washington University, St. Louis, MO (US); and The Arizona Board of Regents for and on Behalf of Arizona State University, Scottsdale, AZ (US)
Filed by The Arizona Board of Regents for and on Behalf of Arizona State University, Scottsdale, AZ (US); and The Washington University, St. Louis, MO (US)
Filed on Oct. 14, 2021, as Appl. No. 17/500,940.
Application 17/500,940 is a continuation of application No. 16/990,033, filed on Aug. 11, 2020, granted, now 11,180,765.
Application 16/990,033 is a continuation of application No. 16/704,032, filed on Dec. 5, 2019, granted, now 10,774,334, issued on Sep. 15, 2020.
Application 16/704,032 is a continuation of application No. 15/850,957, filed on Dec. 21, 2017, abandoned.
Application 15/850,957 is a continuation of application No. 15/040,005, filed on Feb. 10, 2016, granted, now 9,885,051, issued on Feb. 6, 2018.
Application 15/040,005 is a continuation of application No. 13/789,665, filed on Mar. 7, 2013, granted, now 9,297,015, issued on Mar. 29, 2016.
Application 13/789,665 is a continuation of application No. 12/615,872, filed on Nov. 10, 2009, granted, now 8,445,254, issued on May 21, 2013.
Application 12/615,872 is a continuation of application No. PCT/US2008/063293, filed on May 9, 2008.
Claims priority of provisional application 60/917,313, filed on May 10, 2007.
Prior Publication US 2022/0090097 A1, Mar. 24, 2022
Int. Cl. C12Q 1/68 (2018.01); A61K 35/74 (2015.01); A61K 39/00 (2006.01); C12N 1/36 (2006.01); C12N 15/74 (2006.01); A61K 35/00 (2006.01)
CPC C12N 15/74 (2013.01) [A61K 35/74 (2013.01); A61K 39/00 (2013.01); C12N 1/36 (2013.01); A61K 2035/11 (2013.01); A61K 2039/522 (2013.01); A61K 2039/523 (2013.01)] 15 Claims
 
1. A recombinant bacterium capable of regulated expression of at least one nucleic acid sequence encoding a protein of interest, wherein the bacterium comprises:
a. at least one chromosomally integrated nucleic acid sequence encoding a repressor operably linked to a regulatable promoter, wherein the nucleic acid sequence encoding the repressor and/or promoter have been modified from the wild-type nucleic acid sequence so as to optimize the expression level of the nucleic acid sequence encoding the repressor,
wherein the repressor is selected from the group consisting of Lacl, C2, and C1,
wherein the regulatable promoter is selected from the group consisting of PrhaBAD, PxylAB, and PxylFGH;
b. a vector comprising at least one nucleic acid sequence encoding a protein of interest operably linked to a promoter regulated by the repressor, such that the expression of the nucleic acid sequence encoding the protein is repressed during in vitro growth of the bacterium, but the bacterium is capable of high level expression of the nucleic acid sequence encoding the protein in a host; and
c. a deletion in pmi, wherein the deletion is Δpmi-2426.