EP0063949A2
Expired / Lapsed
EP
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Plasmids coding for B. Thuringiensis crystal protein, their production and their use in creating genetically engineered bacterial strains, the aforesaid strains per se and their use in producing said protein, said protein so-produced, insecticidal formulations comprising the same and a method of producing an insecticidal effect
Assignee: Washington Research Foundation
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Filing: 1982-04-26
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Granted: —
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Expires: 2002-04-26
Abstract
Abstract The crystal protein of Bacillus thuringiensis is an insecticidal substance. Unfortunately growth phase limitations are exhibited by B. thuringiensis in producing this protein. This invention, however, provides a plasmid capable of replication in a bacterial host species and containing expressible heterologous DNA coding for the crystal protein of Bacillus thuringiensis and including an expression mechanism for said heterologous DNA which is recognized by the host species system but does not exhibit substantial growth phase limitations in the bacterial host species. Genetically engineered bacterial host strains transformed by the plasmids of the invention express B. thuringiensis crystal proteins without exhibiting the aforesaid growth phase limitations.
Inventors
- 👤 Harry Ernest Schnepf
- 👤 Helen Riaboff Whiteley
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-25 10:47
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/195; C07K14/32; C07K14/325; A; A01; A01N; A01N63/00; A01N63/50; C12; C12N; C12N1/00; C12N1/20; C12N1/205; C12R; C12R2001/00; C12R2001/01; C12R2001/07; C12R2001/075
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
53 words
A plasmid capable of replication in a bacterial host species and containing expressible heterologous DNA coding for the crystal protein of Bacillus thuringiensis and including an expression mechanism for said heterologous DNA which is recognized by the host species system but does not exhibit substantial growth phase limitations in the bacterial host species.
Claim 2
Dependent Claim
53 words
A plasmid capable of replication in a bacterial host species and containing expressible heterologous DNA coding for the crystal protein of Bacillus thuringiensis and including an expression mechanism for said heterologous DNA which is recognized by the host species system but does not exhibit substantial growth phase limitations in the bacterial host species.
Claim 3
Dependent Claim
25 words
A plasmid as claimed in claim 1 comprising a DNA portion having substantial sequence homology to the DNA coding for the crystal protein of B.thuringiensis.
Claim 4
Dependent Claim
32 words
A plasmid as claimed in claim 1 comprising a DNA portion having substantial sequence homology to plasmids of B. thurinaiensis having a molecular mass greater than 1 0 x 10 6 Mr.
Claim 5
Dependent Claim
32 words
A plasmid as claimed in claim 3, wherein said DNA portion is identified with a Pvu II-C DNA fragment probe or a probe having substantial sequence homology with a Pvu II-C probe.
Claim 6
Dependent Claim
60 words
5- A plasmid as claimed in claim 4, wherein said DNA portion is derived from B.thuringiensis subspecies tolworthi; subspecies darmstadiensis; subspecies sotto; subspecies thuringiensis; subspecies thuringiensis, HD-290; subspecies thuringiensis, H D -120; subspecies thuringiensis, HD-2; subspecies kurstaki, HD-244; subspecies kurstaki, HD-73; subspecies kurstaki, HD-1; subspecies alesti, HD-4; subspecies toumanoffi, F-9; subspecies galleriae, HD-8; subspecies wuhnanesis, F-6; or subspecies morrisoni, F-5.
Claim 7
Dependent Claim
27 words
A plasmid according to claim 1 as claimed in any one of claims 1 to 5 including a DNA portion having substantial sequence homology to plasmid pBR322.
Claim 8
Dependent Claim
44 words
The plasmids pESl (ATCC Number 31995), pJWK20 (ATCC Number 31997), and pJWK18 (ATCC Number 31998), a mutant plasmid having the B. thuringiensis crystal protein coding characteristics of said plasmid pESl, or pJWK20, or pJWK18, and progeny of any of said plasmids or mutant plasmids.
Claim 9
Dependent Claim
44 words
The plasmids pESl (ATCC Number 31995), pJWK20 (ATCC Number 31997), and pJWK18 (ATCC Number 31998), a mutant plasmid having the B. thuringiensis crystal protein coding characteristics of said plasmid pESl, or pJWK20, or pJWK18, and progeny of any of said plasmids or mutant plasmids.
Claim 10
Dependent Claim
29 words
A bacterial strain containing a plasmid as claimed in any one of claims 1 to 6 or a plasmid, mutant plasmid or progeny thereof as claimed in claim 7.
Claim 11
Dependent Claim
27 words
The bacterial strains ES12, JWK1, and JWKll, in each case transformed by a plasmid as claimed in claim 1, and B. thuringiensis crystal protein antigen-producing mutants thereof.
Claim 12
Dependent Claim
82 words
A genetically engineered bacterial strain which has been transformed to introduce thereinto heterologous DNA which, upon expression, results in a polypeptide of 130,000 M r which reacts specifically with antibody for the crystal protein of B. thuringierisis. host other than B. thuringiensis, the expression mechanism being recognized by said host without exhibiting substantial growth phase limitations, and ligating said first DNA fragment and said other fragment(s) in the correct arrangement for expression of the hetexologous crystal protein-producing DNA coding in said host.
Claim 13
Dependent Claim
82 words
A genetically engineered bacterial strain which has been transformed to introduce thereinto heterologous DNA which, upon expression, results in a polypeptide of 130,000 M r which reacts specifically with antibody for the crystal protein of B. thuringierisis. host other than B. thuringiensis, the expression mechanism being recognized by said host without exhibiting substantial growth phase limitations, and ligating said first DNA fragment and said other fragment(s) in the correct arrangement for expression of the hetexologous crystal protein-producing DNA coding in said host.
Claim 14
Dependent Claim
77 words
A method of creating a genetically engineered bacterial strain which is capable of producing the crystal protein of Bacillus thuringiensis comprising transforming a host bacterial strain other than B. thuringiensis by introducing thereinto a plasmid as claimed in any one of claims 1 to 6, or a plasmid, mutant plasmid or progeny thereof as claimed in claim 7, whereby the expressible genetic material in the host includes. heterologous DNA coding for the crystal protein of B. thuringiensis.
Claim 15
Dependent Claim
13 words
A method as claimed in claim 15, wherein the host is E. coli.
Claim 16
Dependent Claim
15 words
A method as claimed in claim 16, wherein the host is ES12, JWK1 or JWK11.
Claim 17
Independent Claim
41 words
A method of preparing Bacillus thuringiensis crystal protein comprising growing a bacterial strain as claimed in any one of claims 8 to 10 or which has been created by a method as claimed in any one of claims 15 to 17.
Claim 18
Dependent Claim
66 words
B. thuringiensis crystal protein which has been produced by a genetically engineered bacterial strain transformed by introduction thereinto of a plasmid as claimed in any one of claims 1 to 6 or a plasmid, mutant plasmid or progeny thereof as claimed in claim 7, or by a bacterial strain as claimed in claim 9 or by a genetically engineered bacterial strain as claimed in claim 10.
Claim 19
Dependent Claim
27 words
An insecticidal formulation or preparation comprising a protein as claimed in claim 11 in a form suitable for insecticidal use, optionally including a diluent, carrier or excipient.
Claim 20
Dependent Claim
62 words
A method of producing an insecticidal effect in an environment which comprises treating the environment with a protein as claimed in claim 11 or with a formulation or preparation as claimed in claim 12 or exposing the environment to growth of a bacterial strain as claimed in claim 9 or growth of a genetically engineered bacterial strain as claimed in claim 10.
Claim 21
Independent Claim
59 words
A method of producing as plasmid as claimed in any of claims 1 to 7 which comprises isolating from a crystal protein-producing strain of Bacillus thuringiensis a first fragment of DNA comprising an expressible DNA coding for the crystal protein, providing one or more other DNA fragments which contribute(s) an expression mechanism for heterologous DNA in a selected bacterial
References Cited (Backward Citations)
No citation records indexed.
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