Bacteria for expressing a polysaccharide depolymerase containing a novel recombinant plasmid
AssigneeMicrolife Technics, Inc.
InventorPeter A. Vandenbergh
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Abstract
A segment of a bacteriophage encoding for a polysaccharide depolymerase which has been cloned and expressed in Escherichia coli is described. In particular, bacteriophage ERA103 was found to consist of five EcoRI fragments labeled: A, 7.5-kb; B, 5.0-kb; C, 2.7-kb; D, 2.1-kb and E, 1.8-kb. Fragment B encodes for the depolymerase and was cloned into the positive-selection vector pOP203(A.sub.2.sup.+), pBR322 and the expression vector pKK223-3. The depolymerase is applied to rosaceous plants to prevent Fireblight caused by Erwinia amylovora by depolymerizing a polysaccharide produced by this bacteria.
Claims27
A recombinant plasmid which can express a polysaccharide depolymerase in bacteria containing the plasmid which comprises:
- (a) a linear segment from bacteriophage ERA 103 deposited as ATCC 39824 Bl which infects Erwinia amylovora to produce the depolymerase and which segment codes for the production of the depolymerase;
- and
- (b) a transformable vector plasmid segment ligated to the linear segment to provide the recombinant plasmid wherein the recombinant plasmid is stable in the bacteria so as to express the depolymerase.
A bacteria which expresses a polysaccharide depolymerase because of a resident recombinant plasmid wherein the recombinant plasmid comprises:
- (a) a linear segment from bacteriophage ERA 103 deposited as ATCC 39824 Bl which infects Erwinia amylovora to produce the depolymerase and which segment codes for the production of the depolymerase;
- and
- (b) a transformable vector plasmid segment ligated to the linear segment to provide the recombinant plasmid wherein the recombinant plasmid is stable in the bacteria so as to express the depolymerase.
A method for producing a polysaccharide depolymerase which comprises:
- (a) providing a bacteria for expressing the polysaccharide depolymerase containing a resident recombinant plasmid with a linear segment from bacteriophage ERA 103 deposited as ATCC 39824 Bl which infects Erwinia amylovora to produce the depolymerase and which segment codes for the production of the depolymerase and with a transformable vector plasmid segment which has been ligated to the linear segment to provide the recombinant plasmid wherein the recombinant plasmid is stable in the bacteria so as to express the depolymerase;
- (b) growing the bacterial in a growth medium to produce the depolymerase;
- and
- (c) removing the depolymerase from the bacteria and growth medium.
Dependencies are inferred from the claim text.