Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1995-06-07
1998-10-13
Ketter, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 698, 435 712, 4351723, 435219, 4352529, 4352534, 4353201, C12P 2104
Patent
active
058210881
ABSTRACT:
A novel system for cloning and expression of genes in gram-positive bacteria. The expression system is based on the finding that many gram-positive bacteria sort proteins to their cell surface through cis-acting N-terminal signal sequences and C-terminal anchor regions. In particular, the cell sorting signals of the streptococcal M6 protein, a well-known surface molecule, are used to construct a gram-positive expression system, designated SPEX (Streptococcal Protein Expression). Expression is achieved by cloning the gene of interest into an appropriate SPEX cassette which is then stably introduced into a bacterial host, such as the human commensal Streptococcus gordonii. Depending on the SPEX vector used, recombinant proteins can be anchored to the cell wall prior to release by specific endoproteolytic cleavage or secreted into the culture medium during bacterial growth. The use of host bacteria lacking extracellular proteases should protect secreted proteins from proteolytic degradation. Several expression vectors in this system also produce specifically-tagged recombinant proteins which allows for a one-step purification of the resulting product.
REFERENCES:
patent: 4965197 (1990-10-01), Liebl et al.
patent: 5013653 (1991-05-01), Huston et al.
patent: 5115102 (1992-05-01), Haymore et al.
patent: 5532142 (1996-07-01), Johnston et al.
patent: 5616686 (1997-04-01), Fischetti et al.
Gianni Pozzi et al., "Delivery and Expression of a Heterologous Antigen on the Surface of Streptococci", Infection and Immunity (May 1992) 60:1902-1907.
Gianni Pozzi et al., "Human T-helper cell Recognition of an Immunodominant Epitope of HIV-1 gp120 Expressed on the Surface Streptococcus gordonii", Vaccine (1994) 12:1071-77.
G. Pozzi, "Expression of M6 Protein Gene of Streptococcus pyogenes in Streptococcus gordonii After Chromosomal Integration and Transcriptional Fusion" Infection and Immunity (1992) 143:449-457.
D. Medaglini et al., "Mucosal and Systemic Immune Responses to a Recombinant Protein Expressed on the Surface of the Oral Commensal Bacterium Streptococcus gordonii After Oral Colonization" (1994) PNAS (1995) 92:6868-72.
Darzins Aldis
Fischetti Vincent A.
Hruby Dennis
Whitehead Stephen
Ketter James
Siga Pharmaceuticals, Inc.
Yucel Irem
LandOfFree
Use of gram-positive bacteria to express recombinant proteins does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Use of gram-positive bacteria to express recombinant proteins, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Use of gram-positive bacteria to express recombinant proteins will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-312451