Heterologous gene expression in lactococcus, and the expression products therefrom

• US 6,221,648 B1
• Filed: 10/27/1994
• Issued: 04/24/2001
• Est. Priority Date: 02/27/1992
• Status: Expired due to Fees

• Current Assignee: Microbial Technics Limited
• Original Assignee: Microbial Technics Limited
• Inventors: Wilson, Peter William, De Villareal, Pamela Norton, Wells, Jeremy Mark, Le Page, Richard William Falla
• Primary Examiner(s): PRIEBE, SCOTT DAVID
• Application Number:
  US08/290,995
• Time in Patent Office: 2,371 Days
• Field of Search: 435/252.3, 435/69.1, 435/471, 514/2, 424/93.4, 424/93.44
• US Class Current: 435/252.3
• CPC Class Codes: A01N 63/23 B. thuringiensis
• CPC Class Codes: A01N 63/50 Isolated enzymes; Isolated proteins peptides A01N37/46
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/325 Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
• CPC Class Codes: C07K 14/33 from Clostridium (G)
• CPC Class Codes: C12N 15/746 for lactic acid bacteria (Streptococcus; Lactococcus; Lactobacillus; Pediococcus; Enterococcus; Leuconostoc; Propionibacterium; Bifidobacterium; Sporolactobacillus)
• CPC Class Codes: C12N 2740/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

Heterologous polypeptides are produced in lactococcus using a T7 or T7-like RNA polymerase gene under the control of an inducible promoter effective in a lactococcus host, and a promoter specific for said polymerase upstream of a coding sequence for the heterologous polypeptide. Thus, the promoter specific for the polymerase directs transcription of the coding sequence selectively as a result of expression of the polymerase. The heterologous polypeptide can be produced at high yield, and can be secreted. The polypeptide within the cell, being biologically active, can be delivered in the encapsulated form, e.g. as a medicament, vaccine or as an environmental pesticide.

Claims

1. Lactococcus lactis host organism transformed with recombinant DNA comprising:

a T7 RNA polymerase gene placed under the control of the lac inducible promoter and the lacR gene, and

a promoter specific for the T7 RNA polymerase upstream of a coding sequence for a desired polypeptide, whereby the promoter specific for said T7 RNA polymerase directs transcription of said coding sequence selectively as a result of expression of said polymerase.

2. Lactococcus lactis host organism transformed with recombinant DNA comprising:

a T7 RNA polymerase gene placed under the control of the lac inducible promoter and the lacr gene;

a promoter specific for the T7 RNA polymerase upstream of a coding sequence for a desired polypeptide, whereby the promoter specific for said T7 RNA polymerase directs transcription of said coding sequence selectively as a result of expression of said polymerase; and

,

a secretory signal sequence, derived from a protein endogenous to Lactococcus lactis operatively linked to said coding sequence for the desired polypeptide, whereby the desired polypeptide is secreted from said host organism.