Transfection of enteric parasites
AssigneeUniversity of Virginia Patent Foundation
InventorsWilliam A. Petri, Jr., R. Randolph Vines, Jay E. Purdy, Barbara J. Mann
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Abstract
The invention provides methods for expressing foreign genes in enteric protozoa. This transfection system was established using a gene ligated to the 5' and 3' flanking DNA regions of a protein-encoding gene from an enteric protozoa. The present invention also provides such transformed enteric protozoa, vaccines produced therefrom and foreign or altered proteins expressed in the same. The ability to introduce and express genes in amebae will now permit both genetic analysis and modification of the virulence of this organism, which remains a serious threat to world health and will facilitate basic research towards the control of this parasite.
Claims26
A method for obtaining an enteric protozoan which expresses foreign genes comprising the steps of:
- transfecting the enteric protozoan Entamoeba histolytica with an expression vector containing (i) at least 157 base pairs of 5' flanking DNA sequence isolated from a protein-encoding gene of the enteric protozoan Entamoeba histolytica which enables expression of the foreign gene in said enteric protozoan and (ii) a gene which is to be expressed operably linked to said flanking DNA sequence, and isolating a transformed enteric protozoan.
A method to express a foreign or altered protein in an enteric parasite comprising the steps of:
- transfecting an enteric protozoan Entamoeba histolytica with an expression vector containing (i) at least 157 base pairs of 5' flanking DNA sequence isolated from a protein-encoding gene of an enteric protozoan and (ii) a foreign or altered gene operably linked to said flanking DNA sequence,
- isolating a transformed enteric protozoan,
- and culturing said transformed enteric protozoan,
- wherein said foreign or altered protein is expressed.
A plasmid comprising (i) at least 157 base pairs of 5' flanking DNA sequence isolated from a protein-encoding gene of an enteric protozoan Entamoeba histolytica, which enables expression of a foreign gene in said enteric protozoan and (ii) said foreign gene operably linked to said flanking DNA sequence.
Plasmid pTCV1.
Plasmid pTCV2.
A transformed enteric protozoan, Entamoeba histolytica, containing a plamid comprising (i) at least 157 base pairs of 5' flanking DNA sequence isolated from a protein-encoding gene of Entamoeba histolytica, which enables expression of a foreign gene in said enteric protozoan and (ii) said foreign gene operably linked to said flanking DNA sequence.
Dependencies are inferred from the claim text.