Abstract: The present invention provides minicircle nucleic acid vector formulations for use in administering to a subject, wherein the minicircle nucleic acid vectors include a polynucleotide of interest, a product hybrid sequence of a unidirectional site-specific recombinase, and are devoid of plasmid backbone bacterial DNA sequences. Also provided are methods of producing the subject formulations as well as methods for administering the minicircle nucleic acid vector formulations to a subject. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications.

Abstract: Disclosed herein are zinc fingers comprising CCHC zinc coordinating residues. Also described are zinc finger proteins and fusion proteins comprising these CCHC zinc fingers as well as polynucleotides encoding these proteins. Methods of using these proteins for gene editing and gene regulation are also described.

Abstract: The present invention generally relates to devices and methods for immobilizing nucleic acids on a substrate. In certain embodiments, devices of the invention include a voltage source, and a substrate coupled to the voltage source, in which hydrophobicity of the substrate changes in response to an applied electric field and a surface of the substrate is coated with a substance that retains nucleic acids.

Abstract: The invention provides a bacterium containing a polynucleotide comprising a nucleic acid encoding a heterologous antigen, as well as fusion protein partners. Also provided are vectors for mediating site-specific recombination and vectors comprising removable antibiotic resistance genes.

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

Abstract: Methods and means are provided for the exact exchange in eukaryotic cells, such as plant cells, of a target DNA sequence for a DNA sequence of interest through homologous recombination, whereby the selectable or screenable marker used during the homologous recombination phase for temporal selection of the gene replacement events can subsequently be removed without leaving a foot-print employing a method for the removal of a selected DNA flanked by two nucleotide sequences in direct repeats.

Abstract: The invention relates to a method for the simultaneous integration of two or more copies of a polynucleotide of interest into the chromosome of a fungal host cell comprising at least two pairs of recognition sequences of a site-specific recombinase, each pair flanking a resident negative selection marker; transformation of the cell with a construct carrying a gene of interest also flanked by the recognition sequences to ensure double-crossover events after transient expression of the recombinase, followed by selection for excision of all negative selection markers from the cell.

Abstract: Methods are provided for engineering novel strand-specific nicking endonucleases by means of an in vivo enrichment of a plasmid library containing a randomly mutagenized restriction endonuclease gene. The plasmids contain adjacent to the gene a cleavable or nickable sequence for cleaving or nicking by the endonuclease product of the gene and a second recognition site for a second endonuclease. The plasmid library is used to transform unmodified host cells. Plasmids from the cultured transformed cells may be analyzed by an in vitro assay for nicking and the nicked plasmids pooled and used to transform host cells. The product is then pooled and the single-stranded specificity of the endonuclease is then determined. The product is either cloned after amplification or identified by use of a selectable marker.

Abstract: The present invention provides methods, nucleic acid constructs, and kits for selectively deleting a region of a nucleic acid sequence. Specifically, it utilizes retargeting retroelements to place site-specific recombination sites at targeted locations in the nucleic acid sequence. The region between the recombination sites is then deleted using a site-specific recombination system.

Abstract: This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

Abstract: The invention relates to recombination systems and methods for eliminating nucleic acid sequences from the chromosomal DNA of eukaryotic organisms, and to transgenic organisms—preferably plants—which comprise these systems or were generated using these methods.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: The present invention relates to a novel aspartokinase derived from a Coryneform bacterium; a DNA encoding the enzyme; a recombinant DNA containing the above DNA; a Coryneform bacterium having the above recombinant DNA or a Coryneform bacterium having the DNA on its chromosome; and a process for producing L-lysine by culturing the above microorganism. Construction has been successfully made of a DNA encoding an aspartokinase freed from concerted feedback inhibition by L-lysine and L-threonine derived from a Corynebacterium and has a nucleotide sequence encoding an amino acid sequence wherein the amino acid residue at position 311 is an amino acid other than Thr in the amino acid sequence shown by SEQ ID NO: 18.

Abstract: A single-copy BAC vector (containing or lacking an insert) is converted in a host cell into a conditional high-copy BAC vector by introducing a conditional origin of replication into the single-copy BAC vector. The conditional ori is introduced by site-specific recombination between the SC BAC vector and a vector that contains the conditional ori. The host cell comprises a recombinase that recognizes a site-specific recombination site on both the BAC vector and the conditional ori vector. In the presence of the recombinase, the conditional ori-containing vector recombines into the BAC vector to produce a high-copy BAC vector that can be conditionally amplified by activating the conditional origin of replication on command.

Abstract: A plasmid vector characterized by comprising a promoter sequence that can be recognized by an RNA polymerase which is not inherent in a host and that controls the expression of desired genes and a replication origin that increases the number of copies under the induction by exogenous factors; methods for expression and isolation of target genes by using the vector; a polypeptide having the activity of an AccIII restriction endonuclease; and a DNA encoding the polypeptide. The invention provides for the first time a plasmid vector which can introduce an exogenous desired gene encoding proteins which are lethal or harmful to hosts into the hosts, a method for efficiently expressing the proteins by using the vector, and also a method for permitting a restriction endonuclease-gene constituting a restriction-modification system to be isolated even in the absence of a modification enzyme gene, which has been difficult in the prior arts.

Abstract: A method is provided for identifying a restriction endonuclease that includes: screening a target DNA sequence for the presence of known methylase sequence motifs, identifying any open reading frames which lie close to the screened methylase sequence motif and assaying the protein products of the open reading frames for restriction endonuclease activity.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: The present invention relates to host cells suitable for expressing genes under the direction of foreign RNA polymerases and to providing very low levels of expression of such genes and RNA polymerases in the absence of induction.

Abstract: A process for site-directed integration of multiple copies of a gene in a mould is provided, which comprises transforming a mould cell containing in its chromosomal DNA a restriction site for a rare-cutting endonuclease, e.g., I-Scel, preferably introduced at a desired locus, e.g., within a selectable marker gene or in the neighborhood thereof, with a piece of DNA comprising multiple copies of at least one expressible gene comprising at least one structural gene encoding a desired protein, surrounded by two DNA fragments homologous to part of the DNA upstream and downstream, and in the neighborhood, of said restriction site, while during the transformation of the mould the presence of the rare-cutting endonuclease is provided, followed by selecting or screening for a mould cell in which the multiple gene copies of said expressible gene are inserted into the chromosomal DNA of the mould.

Abstract: Transgenic fluorescent Pseudomonas spp. are described which have a biosynthetic locus which encodes for the production of the antibiotic phenazine-1-carboxylic acid stably introduced into the genome, have a locus which encodes for the production of the antibiotic 2,4-diacetylphloroglucinol, and are effective for control of diseases caused by the soil-borne fungus, Rhizoctonia. Strains are also described which control diseases caused by Gaeumannomyces graminis or Pythium, in addition to Rhizoctonia, or have the ability to control all three diseases.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: SPHINGLY polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing SPHINGLY polypeptides and polynucleotides in therapy, and diagnostic assays for such.

Abstract: A plasmid vector characterized by comprising a promoter sequence that can be recognized by an RNA polymerase which is not inherent in a host and that controls the expression of desired genes and a replication origin that increases the number of copies under the induction by exogenous factors; methods for expression and isolation of target genes by using the vector; a polypeptide having the activity of an AccIII restriction endonuclease; and a DNA encoding the polypeptide. The invention provides for the first time a plasmid vector which can introduce an exogenous desired gene encoding proteins which are lethal or harmful to hosts into the hosts, a method for efficiently expressing the proteins by using the vector, and also a method for permitting a restriction endonuclease gene constituting a restriction-modification system to be isolated even in the absence of a modification enzyme gene, which has been difficult in the prior arts.

Abstract: The present invention relates to the recombinant DNA which encodes the DraIII restriction endonuclease modification methylase, and the production of DraIII restriction endonuclease from the recombinant DNA. Related expression vectors, pHKUV5 vector which features a strong, constitutive UV5 promoter without the Lac repressor binding site and pHKT7 vector which contains a powerful controllable T7 promoter and a low copy number origin of replication, are also disclosed.

Abstract: The present invention relates to the recombinant DNA which encodes the SgrAI restriction endonuclease and the MspI modification methylase, and the production of SgrAI restriction endonuclease from the recombinant DNA.

Abstract: The present invention relates to isolated nucleic acid sequences encoding polypeptides having tripeptide aminopeptidase activity. The invention also relates; to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as recombinant methods for producing the polypeptides.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.