Abstract: The present invention relates to cloning recombinant DNA molecules encoding a multi-specific methylase gene (bssHIIM1), BssHII restriction endonuclease gene (bssHIIR), and the cognate BssHII methylase gene (bssHIIM2) from Bacillus stearothermophilus H3 E. coli. The BssHII multi-specific methylase gene was first cloned in a Sau3AI library using a modified pLITMUS28 vector (New England Biolabs, Inc., Beverly, Mass.) with two BssHII sites. Expression of the multi-specific BssHII methylase renders the two BssHII sites resistant to BssHII digestion. Surprisingly, the cloned methylase also modifies some other sites in addition to BssHII site (5'GCGCGC3'). The methylase also modifies BsrFI site (5'RCCGGY3') and HaeII site (5'RGCGCY3'); and partially modifies EagI site (5'CGGCCG3') and MIul site (5'ACGCGT3'). The beginning of the bssHIIR gene was cloned by using two degenerate primers based on the N-terminal amino acid sequence in PCR. The rest of the bssHIIR gene was cloned by inverse PCR.

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: 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: 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 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 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 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 provides a novel approach to the production of restriction endonucleases. More specifically, there is provided a novel method for the overexpression of these enzymes, which comprises transforming a host cell with an expression vector containing DNA coding for the restriction endonuclease of interest and a vector for the expression of a methylase which is capable of protecting the host cell DNA against the restriction endonuclease of interest. The methylase vector is compatible with the restriction endonuclease expression vector. Preferably, the expression vector is a T7 expression vector and the cell is transformed with a third compatible vector which regulates the expression vector. Also disclosed is the cloning and overexpression of the AatII restriction endonuclease and methylase and the cloning and overexpression of the AluI restriction endonuclease and methylase.

Abstract: A process for preparing silver-containing catalysts and their carriers for the production of ethylene oxide via ethylene oxidation and also to the applications of said catalysts in producing ethylene oxide.Commercial trihydrated .alpha.-alumina, boehmite, carbonaceous materials, a fluxing agent, fluoride and a binder are mixed with water, kneaded and extruded to form strips which are cut and shaped. The shaped bodies are dried, calcined and converted to a .alpha.-alumina bodies i.e., carriers. This process is characterized by using trihydrated .alpha.-alumina, boehmite alumina and carbonaceous materials which have a good matching of particle sizes and proportions in preparing alumina carriers with the following pore structure:______________________________________ specific surface area 0.2-2 m.sup.2 /g pore volume >0.5 ml/g pore radius >30.mu., 25-10% of total volume <30.mu., 75-90% of total volume.