Abstract
Expression of the crystal protein of Bacillus thuringiensis in Escherichia coli is described by use of plasmids containing heterologous DNA coding for the crystal protein. Genetically engineered bacterial host strains transformed by the plasmids of the invention express Bacillus thuringiensis crystal proteins without exhibiting the growth phase limitations characteristic of the natural bacterial host species.
Filing date: Mar 30, 1982
Issue date: Aug 21, 1984
Inventors: H. Ernest Schnepf, Helen R. Whiteley
Assignee: The Board of Regents of the University of Washington
Primary Examiner: James Martinell
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What is claimed is:
1. A hybrid recombinant plasmid capable of replication in an Escherichia coli bacterial host species, said plasmid containing expressible heterologous DNA coding for a polypeptide which has the immunological properties of the crystal protein of Bacillus thuringiensis, said plasmid further containing expressible heterologous DNA having a DNA portion derived from plasmids of Bacillus thuringiensis having a molecular mass greater than 10.times.10.sup.6 M.sub.r, said Bacillus thuringiensis derived DNA portion being identifiable with a Pvu II-C DNA fragment probe, said hybrid recombinant plasmid further comprising an expression mechanism for said expressible heterologous DNA which is recognized by the host species' system.
2. A hybrid recombinant plasmid capable of replication in an Escherichia coli bacterial host species, said plasmid containing expressible heterologous DNA coding for a polypeptide which has the immunological properties of the crystal protein of Bacillus thuringiensis, said plasmid further containing expressible heterologous DNA having a DNA portion derived from plasmids of Bacillus thuringiensis having a molecular mass greater than 10.times.10.sup.6 M.sub.r, said Bacillus thuringiensis derived DNA portion further being identifiable with a Pvu II-C DNA fragment probe, said Bacillus thuringiensis derived DNA portion further being selected from the group consisting of Bacillus thuringiensis subspecies tolworthi; Bacillus thuringiensis subspecies darmstadiesis; Bacillus thuringiensis subspecies sotto; Bacillus thuringiensis subspecies thuringiensis; Bacillus thuringiensis subspecies thuringiensis, strain HD-290; Bacillus thuringiensis subspecies thuringiensis, strain HD-120; Bacillus thuringiensis subspecies thuringiensis, strain HD-2; Bacillus thuringiensis subspecies kurstaki, strain HD-244; Bacillus thuringiensis subspecies kurstaki, strain HD-73; Bacillus thuringiensis subspecies kurstaki, strain HD-1; Bacillus thuringiensis subspecies alesti, strain HD-4; Bacillus thuringiensis subspecies toumanoffi, strain F-9; Bacillus thuringiensis subspecies galleriae, strain HD-8; Bacillus thuringiensis subspecies wuhnanesis, strain F-6 and Bacillus thuringiensis subspecies morrisoni, strain F-5; said hybrid recombinant plasmid further including an expression mechanism for said heterologous DNA which is recognized by the host species' system.
3. Hybrid recombinant plasmids having the Bacillus thuringiensis crystal protein coding characteristics of plasmids: pES1, as carried in Escherichia coli strain ES12, (ATCC Number 31995); pJWK20, as carried in Escherichia coli strain JWKI (ATCC Number 31997); and pJWK18, as carried in Escherichia coli strain JWKII, (ATCC Number 31998), and their progeny resulting from normal cell division of the parental bacterial cells or plasmids.