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Amerithrax — Part 27
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United States Patent: 6,316,006 @ r Page 2 of 10
Primary Examiner: Housel; James C.
Assistant Examiner: Shaver; Jennifer
Attorney, Agent or Firm: Harris; Charles H., Moran; John Francis
Claims
What is claimed is:
1. A recombinant asporogenic B. anthracis isolated from .DELTA.Sterne-1 (pPA102) which shows
inability to bind the dye when grown on Congo Red Agar.
2, AB. anthracis of claim 1 which is B. anthracis DELTA.Sterne-1(pPA102)CR4.
3. A composition comprising the organism of claim 1 in a growth medium.
A, A composition comprising the organism of claim 2 in a growth medium.
Description
FIELD OF THE INVENTION
This invention relates to the bacterial expression system, production and use of protective antigen (PA)
against Bacillus anthracis. The PA immunogen is useful in vaccine against human anthrax. The PA can
be produced by an asporogenic organism which overproduces the desired antigen, which is then
harvested from the supernatant.
BACKGROUND OF THE INVENTION
Bacillus anthracis is the etiologic agent responsible for anthrax, a disease often found in persons
exposed to infected animals or their products. Persons particularly exposed to animals include
veterinarians, laboratory technicians, ranchers and employees working with skin or hair of animals. The
mode of entry into the body may be the skin or, when contaminated meat is eaten, the gastrointestinal
tract. Inhaling of spores can cause inhalation anthrax, a disease that can be fatal. Vaccines against
Bacillus anthracis have been available. Virulent strains of the organism produce two toxins and a poly-
D-glutamic acid capsule which are coded for on two endogenous plasmids, pX01 and pX02,
respectively. Loss of either of the plasmids results in an attenuated strain of reduced virulence, while
loss of both results in an avirulent organism. The history of the USAMRID Sterne strain of B. anthracis
prior to 1981 is uncertain, though it is believed to be derived from the Sterne strain isolated at the
Onderstpoort Research Laboratory in Pretoria, South Africa.
In 1985 the Bacillus anthracis protective antigen (PA) gene was cloned into a plasmid (pUB110)
resulting in the formation of a recombinant plasmid identified as pPA102, which was reported in the
literature Zvins and Welkos, Infection and Immunity, 854:537-542 (1986)). The production of vaccines
lacking lethal factor was possible thereby. However, a primary problem remained, since the Bacillus
anthracis formed spores. Once spores have formed, they persist in the environment for months and
years. Once the laboratory environment contains such spores, it is very difficult to free the environment
http://patft.uspto.gov/netacgi/nph-Parser?Sectl=PTO2&Sect2=HITOFF &u=/netahtml/sear... 6/28/2005
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