Non-hemolytic clya for excretion of proteins
AssigneeUniversity of Maryland, Baltimore
InventorsJames E. Galen, Yuansha Chen
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Abstract
The disclosure below provides a protein export system utilizing non-hemolytic variants of HlyE family member proteins for efficiently producing recombinant protein from a host cell. In a preferred embodiment, the protein export system utilizes protein export machinery endogenous to the host bacterium into which the protein export system vector is introduced.
Claims20
A method for producing a fusion protein, comprising:
- (a) transforming a population of bacteria with an expression vector encoding a fusion protein, wherein said fusion protein comprises a protein of interest linked to the carboxy terminus of an export protein, wherein said export protein is a Salmonella enterica serovar Typhi (S. Typhi) cytolysin A (ClyA) protein having substantially reduced hemolytic activity, and
- (b) culturing transformed bacteria of (a) in a culture medium under conditions such that said fusion protein is expressed and exported into the culture medium.
A method for eliciting an immune response to a fusion protein in a subject comprising:
- administering to a subject a population of bacteria which produces and exports a fusion protein in an amount sufficient to elicit an immune response in said subject to said fusion protein, wherein said bacteria comprise an expression vector encoding said fusion protein, wherein said fusion protein comprises a protein of interest linked to the carboxy terminus of an export protein, and wherein said export protein is a Salmonella enterica serovar Typhi (S. Typhi) cytolysin A (ClyA) protein having substantially reduced hemolytic activity, thereby eliciting an immune response to said fusion protein in said subject.
An expression vector comprising an expression cassette, wherein the expression cassette comprises an export protein coding sequence linked to a protein of interest coding sequence in a 5′ to 3′ arrangement, wherein said export protein is a Salmonella enterica serovar Typhi (S. Typhi) cytolysin A (ClyA) protein having substantially reduced hemolytic activity.
Dependencies are inferred from the claim text.