Abstract: The present invention relates to a method for cloning and expressing an exogenous phosphorylation-dependent protein kinase in E. coli, and in particular to the co-expression of one or more components of a signal-transduction pathway in E. coli.

Abstract: The present invention is directed to an extremely thermostable enzyme. More specifically, the invention is directed to a thermostable DNA polymerase 9.degree.N-7, as well as the recombinant form of 9.degree.N-7. In another embodiment, there is provided a method for mutagenizing the exo motif I of the 3'-5' exonuclease DNA polymerases from the native conserved DXE in the 3'-5' exo motif I DNA segment to DXD or AXA.

Abstract: The present invention is directed to a method for cloning and producing the HphI restriction endonuclease by 1) linking up the restriction gene to transcription elements that can be processed in E. coli and 2) expressing DNA modification enzymes in the host that protect against HphI digestion.

Abstract: The present invention relates to isolated DNA coding for the restriction endonuclease SCaI as well as to a method for cloning methylase genes from Streptomyces into E. coli by a modification of the methylase selection method. At first, the standard methylase gene selection method was tried to clone the SCaI methylase gene using a high-copy-number cloning vector pUC19 during library construction. The SCaI methylase gene was refractory to cloning by using pUC19, presumably due to the poor expression of the SCaI methylase gene in E. coli. If the SCaI methylase is not efficiently expressed in E. coli, the SCaI sites on the plasmid will not be sufficiently modified by the methylase. As a consequence, the plasmid will be cleaved and lost in the plasmid library after SCaI endonuclease challenge. Since the standard methylase selection did not work, the "endo-blue method" was tried to clone the SCaI endonuclease gene.

Abstract: Multipurpose cloning in vitro phagemid vectors are disclosed, which vectors may be used to generate high specific-activity RNA probes. The multipurpose vectors also possess a bidirectional in vitro transcription system. Methods for constructing this system are also disclosed.

Abstract: The present invention relates to recombinant DNA which encodes the SapI restriction endonuclease and modification methylase, and the production of SapI restriction endonuclease from the recombinant DNA as well as to methods for cloning Actinomycetes genes into suitable hosts such as E. coli.

Abstract: Methods and products are provided for producing and/or purifying virtually any hybrid polypeptide molecule employing recombinant DNA techniques. More specifically, a DNA fragment coding for a protein molecule, e.g. a polypeptide or portion thereof, is fused to a DNA fragment coding for a binding protein, such as the gene coding for the maltose binding protein. The fused DNA is inserted into a cloning vector and an appropriate host transformed. Upon expression, a hybrid polypeptide is produced which can be purified by contacting the hybrid polypeptide with a ligand or substrate to which the binding protein has specific affinity, e.g. by affinity chromatography. The hybrid polypeptide so purified may in certain instances be useful in its hybrid form, or it may be cleaved to obtain the protein molecule itself by, for example, linking the DNA fragments coding for the target and binding proteins with a DNA segment which codes for a peptide which is recognized and cut by a proteolytic enzyme, such as Factor Xa.

Abstract: The present invention is directed to a method for cloning and producing the SfiI restriction endonuclease by 1) introducing the restriction endonuclease gene from S. fimbriatus into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the SfiI restriction endonuclease activity; and 3) purifying the SfiI restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the SfiI restriction endonuclease activity.

Abstract: The present invention relates to a method of cloning ApaLI methylase gene (apaLIM) and ApaLI endonuclease gene (apaLIR) from Acetobacter pasteurianus into E. coli by the methylase selection method and inverse PCR. The ApaLI methylase gene was cloned into pUC19 (3 ApaLI sites inserted) by the methylase selection method. Eight ApaLI-resistant clones were isolated and found to contain apaLIM gene. However, these clones are not stable such that sometimes overnight cultures were lysed or plasmid DNA was lost in the unlysed culture.

Abstract: The present invention is directed to a method for cloning and producing the FseI restriction endonuclease by 1) introducing the restriction endonuclease gene from Frankia species into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the FseI restriction endonuclease activity, and 3) purifying the FseI restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the FseI restriction endonuclease activity.

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

Abstract: The present invention is directed to a method for cloning and producing the SspI restriction endonuclease by 1) introducing the restriction endonuclease gene from Sphaerotilus species into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the vector encoding and expressing the SspI restriction endonuclease, and 3) purifying the SspI restriction endonuclease from the fermented host which contains the vector encoding and expressing the SspI restriction endonuclease activity.

Abstract: Recombinant DNA polymerases from archaebacteria as well as isolated DNA coding for such polymerases are provided. The isolated DNA is obtained by use of DNA or antibody probes prepared from the DNA encoding T. litoralis DNA polymerase and the T. litoralis DNA polymerase respectively. Also provided are methods for producing recombinant archaebacteria thermostable DNA polymerase and methods for enhancing the expression of such polymerases by identifying, locating and removing introns from within the DNA coding for such DNA polymerases.

Abstract: The present invention discloses a novel method for the direct cloning of nuclease genes such as restriction endonuclease genes in E. coli. In addition, there is provided a novel strain which facilitates application of the method. This method has been successfully employed to clone a number of genes coding for endonuclease including restriction endonuclease genes.

Abstract: The present invention is directed to modified proteins and methods of their production. The modified proteins comprise a controllable intervening protein sequence (CIVPS) inserted into a target protein, the CIVPS being capable of excision from the modified protein under predetermined conditions, i.e., increase in temperature, exposure to light, unblocking of amino acid residues by dephosphorylation or deglycosylation. If desired, the modified protein can be subjected to these conditions. The CIVPS may also be inserted into a region that substantially inactivates target protein activity.

Abstract: The present invention discloses a novel method for the direct cloning of nuclease genes such as restriction endonuclease genes in E. coli. In addition, there is provided a novel strain which facilitates application of the method. This method has been successfully employed to clone a number of genes coding for endonuclease including restriction endonuclease genes.

Abstract: The present invention relates to the use of these cyclophilins, hereinafter referred to as "cyclophilin-like proteins (CLP)", in a method for identifying compounds capable of binding to and/or inhibiting the enzymatic activity of these proteins. Such compounds may be further screened for their ability to inhibit parasites which are not susceptible to the anti-parasitic effects of CsA.

Abstract: The present invention is directed to a method for cloning and producing the BglII endonuclease by: 1) introducing the restriction endonuclease gene from Bacillus globigii into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the BglII restriction endonuclease activity; and 3) purifying the BglII restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the BglII restriction endonuclease.

Abstract: The present invention relates to a recombinant McrBC endonuclease obtainable from Escherichia coli, two components of which, McrB.sub.L and McrC, have been purified in active form. McrBC is active in the presence of GTP and at a low pH. The McrBC endonuclease is also substantially free of a third component, McrB.sub.S, which is believed to inhibit or otherwise interfere with the activity of the enzyme. McrBC has various desirable properties, including the ability to recognize a methylated DNA sequence and also its ability to cleave such a sequence in the presence of GTP. Also provided is a process for the production of recombinant McrBC endonuclease, a process for the determination of the modification state of DNA a process for the determination of an epigenetic alteration or defect (including "imprinting"), as well as a process for identifying and isolating additional enzymes that cleave modified DNA.

Abstract: The present invention is directed to a method for cloning and producing the Hinc II restriction endonuclease by (1) introducing the restriction endonuclease gene from Haemophilus influenzae Rc into a host whereby the restriction gene is expressed; (2) fermenting the host which contains the vector encoding and expressing the Hinc II restriction endonuclease, and (3) purifying the Hinc II restriction endonuclease from the fermented host which contains the vector encoding and expressing the Hinc II restriction endonuclease activity.