IP Library › Granted Patent US 10,774,390
Granted Patent B2
US 10,774,390 · App. 15/754,288 · Granted Sep 15, 2020

Methods and compositions for modulating PrfA mediated virulence gene activation in

Inventors: Daniel A. Portnoy (Albany, CA); Jonathan L. Portman (Berkeley, CA)
Assignee: The Regents of the University of California
C12Q1/689A61K35/74A61K39/0208A61P31/04C07K14/195C12N1/20C12N15/74A61K2039/521C12Q2600/158
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Quick Facts
Patent No.
US 10,774,390
App. No.
15/754,288
Granted
Sep 15, 2020
Kind
B2
Abstract

Methods of assessing expression of a PrfA mediated expression construct in a Listerial host cell are provided. Aspects of the methods include maintaining a Listerial host cell that includes a PrfA mediated expression construct in a defined medium containing a reducing agent and evaluating expression of a product from the expression construct to assess expression of the PrfA mediated expression construct. Also provided are methods of producing and using a Listerial vaccine, as well as defined media for use in such methods.

Claims (12)

1. A method of assessing expression of a PrfA mediated expression construct, the method comprising:

maintaining a Listerial host cell comprising a PrfA mediated expression construct in a defined medium containing a PrfA activating amount of a reducing agent; and

evaluating expression of a product from the expression construct to assess expression of the PrfA mediated expression construct.

2. The method according to claim 1 , wherein the PrfA mediated expression construct is a modified virulence factor expression construct.

3. The method according to claim 1 , wherein the modified virulence factor expression construct is an ActA expression construct.

4. The method according to claim 3 , wherein the ActA expression construct comprises the ActA promoter.

5. The method according to claim 4 , wherein the ActA expression construction encodes a fusion protein.

6. The method according to claim 5 , wherein the fusion protein comprises an N-terminal ActA domain.

7. The method according to claim 1 , wherein the PrfA mediated expression construct encodes a heterologous antigen.

8. The method according to claim 1 , wherein the assessing comprises comparing an observed expression level to a threshold value.

9. The method according to claim 1 , wherein the Listerial host cell is Listeria monocytogenes.

10. The method according to claim 1 , wherein the defined medium is not a brain heart infusion medium or a tryptic soy broth medium.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 21, 2018
From: PORTNOY, DANIEL A.; PORTMAN, JONATHAN L.
To: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Reel/Frame 046201/0350 →
CONFIRMATORY LICENSE Recorded Mar 30, 2018
From: UNIVERSITY OF CALIFORNIA BERKELEY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 045801/0277 →
Continuity (2)
Provisional Application 62213579 · Sep 2, 2015
Related Publication 20180237828A1 · Aug 23, 2018