Claims
- 1. A transformed bacterium for use as a production cell, and having:
(a) a lipooligosaccharide or lipopolysaccharide having a core region containing a terminal heptose; (b) an enzyme capable of adding an acceptor molecule to the terminal heptose; and (c) an inserted sequence of isolated DNA encoding glycotransferase forelongation of the acceptor molecule by sequential addition of saccharide monomer to produce ologosaccharide or polysaccharide.
- 2. The bacterium of claim 1, which is Gram-negative.
- 3. The bacterium of claim 2, which contains an rfe gene.
- 4. The bacterium of claim 3, which contains an lsgG regulatory gene.
- 5. The bacterium of claim 2, which is Escherichia coli or Salmonella minnesota.
- 6. The bacterium of claim 2, which is Escherichia coli K12 strain JM 109.
- 7. The bacterium claim 1, which has terminal heptose on a kdo core.
- 8. The bacterium of claim 1, wherein the enzyme is capable of adding an acceptor molecule which is N-acetyl glucosamine.
- 9. The bacterium of claim 1, having inserted an isolated DNA sequence encoding a glycotransferase catalysing the synthesis of an oligosaccharide of Haemophilus influenzae, Neisseria spp. or Salmonella spp.
- 10. A process for producing a complex carbohydrate, which comprises the steps of:
(a) inoculating production cells which are bacteria according to any of claims 1-9 into a culture medium capable of supporting the growth of said production cells; (b) allowing the growth of said production cells; and (c) recovering the complex carbohydrate from the culture medium.
- 11. The process of claim 10, which comprises the steps of extracting chimeric lipoologosaccharide or lipopolysaccharide from the cultured cells, hydrolysing the extracted lipoologosaccharide or lipopolysaccharide, and recovering the resulting ologosaccharide or polysaccharide.
- 12. The process of claim 10, wherein the ologosaccharide or polysaccharide is incorporated into a vaccine against a Gram-negative pathogen.
CLAIM OF PRIORITY
[0001] This application is a continuation under 35 U.S.C. 111(a) of International Application No. PCT/US00/13538 filed May 18, 2000 and published as WO 00/70060 on Nov. 23, 2000, which claims priority under 35 U.S.C. 119(e) from U.S. Provisional Application Serial No. 60/134,756, filed May 18, 1999, which applications are is incorporated herein by reference.
GOVERNMENTAL RIGHTS
[0002] The United States Government retains certain rights in this invention. Financial support was provided by the National Institute of Allergy and Infectious Diseases under Grant Number A124016 and from the NIH National Center for Research Resources under Grants Number RR01614 and RR04112.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60134756 |
May 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
PCT/US00/13538 |
May 2000 |
US |
Child |
09991374 |
Nov 2001 |
US |