Abstract: The invention provides vectors and methods for directional cloning.

Abstract: The present invention provides methods and compositions related to modulating the resistance of a cell against a target nucleic acid or a transcription product thereof. In some preferred embodiments, the present invention provides compositions and methods for the use of one or more cas genes or proteins for modulating the resistance of a cell against a target nucleic acid or a transcription product thereof. In some embodiments, the present invention provides methods and compositions that find use in the development and use of strain combinations and starter culture rotations. In additional embodiments, the present invention provides methods for labelling and/or identifying bacteria. In some preferred embodiments, the present invention provides methods for the use of CRISPR loci to determine the potential virulence of a phage against a cell and the use of CRISPR-cas to modulate the genetic sequence of a phage for increased virulence level.

Abstract: The present invention relates to plasmid curing, and particularly to efficient and stress-free methods for displacing resident or endogenous plasmids from a host cell, such as a bacterium. The invention extends to method of displacing a plasmid comprising a post-segregational killing system from a host cell, the method comprising introducing a recombinant nucleic acid molecule into a host cell harboring a plasmid comprising a post-segregational killing (PSK) system, characterized in that the recombinant nucleic acid molecule is adapted to neutralize the toxic effects of the plasmid's post-segregational killing system, and wherein the nucleic acid molecule is also adapted to outcompete or inhibit replication of the plasmid. The invention further extends to recombinant nucleic acid molecules that can be used in this method, as well as further uses of the methods and nucleic acid molecules of the invention.

Abstract: The present invention provides novel C. botulinum conjugatively transmissible plasmids and methods of use thereof. Specifically, described herein are novel, conjugatively transmissible clostridial plasmids which are capable of being transferred among and between clostridial species. The novel plasmids of the present invention therefore permits the delivery of heterologous clostridial genes into a clostridial host, such as C. botulinum, and the expression of genes of interest in that host, including clostridial toxins and the nontoxigenic components of the toxin complex, toxin fragments, or antigenic portions thereof, in a way both that ensures abundant expression and facilitates purification. Furthermore, toxins with altered structures, chimeric, hybrid toxins, and other toxin derivatives valuable in medicine could be synthesized in this system.

Abstract: The present invention is related to a reversible, parallel and/or multitask cloning method and kit, which improve the cloning of (preferably multiple) genetic element(s) in a nucleic acid construct such as vector or in chromosome of a cell and the rapid and efficient selection of said construct with a correct integration of said genetic element(s) either in vitro or in vivo.

Abstract: The present invention provides novel C. botulinum conjugatively transmissible plasmids and methods of use thereof. Specifically, described herein are novel, conjugatively transmissible clostridial plasmids which are capable of being transferred among and between clostridial species. The novel plasmids of the present invention therefore permits the delivery of heterologous clostridial genes into a clostridial host, such as C. botulinum, and the expression of genes of interest in that host, including clostridial toxins and the nontoxigenic components of the toxin complex, toxin fragments, or antigenic portions thereof, in a way both that ensures abundant expression and facilitates purification. Furthermore, toxins with altered structures, chimeric, hybrid toxins, and other toxin derivatives valuable in medicine could be synthesized in this system.

Abstract: Compositions and methods are provided for expression of a toxic protein in a host cell preferably a bacterial host cell where at least one T7 RNA polymerase gene Is contained on the host cell chromosome and one or more genes encoding a T7 RNA polymerase inhibitor is located on an F? plasmid or on the chromosome.

Abstract: The invention relates to a system for stable maintenance of a plasmid, to host cells for use in this system and to methods of using the system to obtain a plasmid useful in medical applications. In particular, the invention provides transformed host cell containing: i) a chromosomal gene which inhibits cell growth; and ii) a plasmid encoding an antisense sequence, wherein the antisense sequence encoded by the plasmid inhibits the action of the chromosomal gene, thereby permitting cell growth and a method for stable maintenance of a plasmid in a host cell in vivo.

Abstract: A cloning and/or sequencing vector enables recombinant clones to be selected directly. The vector encodes a fusion protein which includes a protein poison.

Abstract: The present invention relates generally to a Plasmid Maintenance System for the stabilization of expression plasmids encoding foreign antigens, and methods for making and using the Plasmid Maintenance System. The invention optimizes the maintenance of expression plasmids at two independent levels by: (1) removing sole dependence on balanced lethal maintenance functions; and (2) incorporating at least one plasmid partition function to prevent random segregation of expression plasmids, thereby enhancing their inheritance and stability. The Plasmid Maintenance System may be employed within a plasmid which has been recombinantly engineered to express a variety of expression products.

Abstract: The present invention relates generally to a Plasmid Maintenance System for the stabilization of expression plasmids encoding foreign antigens, and methods for making and using the Plasmid Maintenance System. The invention optimizes the maintenance of expression plasmids at two independent levels by: (1) removing sole dependence on balanced lethal maintenance functions; and (2) incorporating at least one plasmid partition function to prevent random segregation of expression plasmids, thereby enhancing their inheritance and stability. The Plasmid Maintenance System may be employed within a plasmid which has been recombinantly engineered to express a variety of expression products.

Abstract: The invention provides methods for identifying a modulator of quorum sensing signaling in bacteria, and for identifying a quorum sensing controlled gene in bacteria. In addition, the invention provides quorum sensing controlled genetic loci in Pseudomas aeruginosa. Novel indicator strains and vectors for engineering the strains for use in the method of the invention are also provided.

Abstract: A method for introducing and expressing genes in animal cells is disclosed comprising infecting the animal cells with live invasive bacteria, wherein the bacteria contain a eukaryotic expression cassette encoding the gene. The gene may encode, e.g., a vaccine antigen, a therapeutic agent, an immunoregulatory agent or an anti-sense RNA or a catalytic RNA.

Abstract: A method for modifying structural gene sequences to enhance the expression of the protein product is disclosed. Also disclosed are novel structural genes which encode insecticidal proteins of B.t.k. HD-1, B.t.k. HD-73, B.t. tenebrionis, B.t. entomocidus, 2 protein of B.t.k. HD-1, and the coat protein of potato leaf roll virus.

Abstract: The invention concerns a new tool for efficient mutagenesis enabling the generation of a collection of mutants in fungi by random insertion of a characterized Fusarium oxysporum Impala transposon in the genome of said fungi. The invention also concerns the resulting mutants.

Abstract: Disclosed and claimed are novel toxins and genes obtainable from Bacillus laterosporus isolates disclosed herein. In preferred embodiments, the subject genes and toxins are used to control Western corn rootworm.

Abstract: A enterotoxin-deficient mutant of a member strain of the Bacillus cereus group does not produce HBL enterotoxin, which has been regarded as a human pathogen found in member strains. An enterotoxin-deficient mutant is suitable for use as a biocontrol agent. Methods for making the mutant and for using the mutant are described.

Abstract: This invention relates to an isolated nucleic acid fragment encoding scorpion toxins that are K-channel modifiers. The invention also relates to the construction of a chimeric gene encoding all or a substantial portion of the K-channel modifier, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the K-channel modifier in a transformed host cell.

Abstract: The present invention relates to a class of microbial coding sequences the transcription or cotranscription of which is specifically induced during microbial infection of a host. These particular coding sequences or defined regions thereof may be used as probes to identify and isolate microbial virulence genes. The products of these virulence genes will provide potential targets for the development of vaccines or antimicrobial agents.

Abstract: The present invention provides methods for eliminating plants containing non-T-DNA sequences derived from a T-DNA vector. More specifically, the present invention provides a method for killing plant cells that receive non-T-DNA sequences based on incorporation of a lethal polynucleotide sequence into the non-T-DNA portion of the vector.

Abstract: The invention provides a biocontrol agent against larvae of mosquitoes and blackflies comprising transgenic Anabaena PCC 7120 carrying a synergistic combination of the endotoxin genes CryIVA and CryIVD of Bacillus thuringiensis subsp. israelensis.

Abstract: Disclosed and claimed are novel toxins and genes obtainable from Bacillus laterosporus isolates disclosed herein. In preferred embodiments, the subject genes and toxins are used to control Western corn rootworm.

Abstract: Methods of preparing recombinant Bacillus anthracis protective antigen or a variant or fragment thereof for use in vaccines is disclosed. The protein is expressed in a recombinant microorganism which comprises a sequence which encodes PA or said variant or fragment thereof wherein either (i) a gene of the microorganism which encodes a catabolic repressor protein and/or AbrB is inactivated, and/or (ii) wherein a region of the PA sequence which can act as a catabolic repressor binding site and/or an AbrB binding site is inactivated. Useful quantities of protein are obtainable from these organisms.

Abstract: New pathogen-induced promoters are provided, particularly nematode-induced promoters, which are characterized by their selective induction of expression in the vicinity of the pathogen infection sites, such as the fixed feeding cells induced by infection of the plant by nematodes. Further provided are chimeric genes comprising these promoters as regulatory elements, as well as transgenic plants, comprising those chimeric genes, which are less susceptible to pathogen infections.

Abstract: A novel tissue-specific promoter is provided which has been isolated from the upstream non-coding region of a plant UFO gene. This promoter, operably associated with a nucleic acid sequence expressing a product of interest, initiates and regulates the transcription of such sequences in a shoot meristem-specific tissue.

Abstract: A cloning and/or sequencing vector enables recombinant clones to be selected directly. The vector encodes a fusion protein which includes a protein poison.

Abstract: A method for introducing and expressing genes in animal cells is disclosed comprising infecting said animal cells with live invasive bacteria, wherein said bacteria contain a eukaryotic expression cassette encoding said gene. The gene may encode, e.g., a vaccine antigen, an therapeutic agent, an immunoregulatory agent or a anti-sense RNA or a catalytic RNA.

Abstract: Cloning systems useful for the isolation of recombinant nucleic acid are disclosed in which the recombination of cloning-system nucleic acid and foreign nucleic acid is linked to the expression of a moiety on the surface of a host organism, the moiety being a first member of a binding pair. When recombination occurs between the nucleic acid and the foreign nucleic acid, the moiety is expressed on the surface of the host organism. The isolation of recombinant nucleic acid is then performed by attaching a second member of the binding pair to a solid support and contacting the host organism with the support. When the first member of the binding pair is expressed on the surface of the host organism, the host organism binds to the second member of the binding pair attached to the solid support, thereby selectively isolating those organisms.

Abstract: The present invention relates to recombinant Yersinia and the use thereof for delivery of proteins into eukaryotic cells, including related compositions and methods of treatment and related assays.

Abstract: The invention relates to a vector for transforming microorganisms capable of undergoing sporulation and which contains itself a heterologous insert comprising a DNA sequence coding for at least part of crystal protein, particularly that of B. thuringensis. It also concerns the polypeptides expressed by said microorganisms and having insecticidal properties similar to those of crystal protein, the microorganisms themselves, as transformed by this vector and insecticidal composition including either said polypeptide or the microorganism itself.

Abstract: Strains of Trichoderma spp. that are improved in their capacity as biological control agents for phytopathogenic fungi and nematodes are obtained that are able to over-produce a proteinase, with the aid of a transformation method which involves introducing the gene prb1 of Trichoderma that codes for the proteinase PrB1 under the control of adequate means of regulating expression, producing multiple copies in a stable manner; this ensures that the control achieved of the disease that is caused by pathogenic fungi or nematodes is better in the transgenic strain than in the uncultivated strain that is used as a receptor of the genetic information.

Abstract: Avirulent Vibrio cholerae strains of O1 (CVD111) and non-O1 (CVD112 and CVD112RM) serogroups having the DNA of the cholera toxin core and the RS1 sequences of the cholera toxin locus deleted, and further having a DNA encoding a resistance to mercury, and a DNA encoding the cholera toxin B subunit, or a part thereof sufficient to confer immunogenicity, re-inserted in the chromosome. Methods of making the avirulent V. cholerae O1 and non-O1 strains of the invention, and cholera vaccines using these strains.

Abstract: A method for introducing and expressing genes in animal cells is disclosed comprising infecting said animal cells with live invasive bacteria, wherein said bacteria contain a eukaryotic expression cassette encoding said gene. The gene may encode, e.g., a vaccine antigen, an therapeutic agent, an immunoregulatory agent or a anti-sense RNA or a catalytic RNA.