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5420009 Detection of xanthomonas campestris pv. citri by hybridization and polymerase chain reaction assays
338
PATENT ABSTRACTS
working under vacuum which remove the ether after the completion of treatment; a gas chromatograph with automatic sampling for determining the residues of ether and the safety of the treated components; and mechanisms for returning the treated and safe components, in conjunction with intravenous fluids, separately or together with the healthy components, to the patient. The treatment cycle is repeated until the patient's blood is made free of virus.
5419899 PURIFIED, R E D U C E D R E C O M B I N A N T INTERLEUKIN-2 COMPOSITIONS Koths Kirston; Thomson James; Kunitani Michael; Wilson Kenneth; Hanisch Wolf Berkeley, CA, 94702, UNITED STATES A process for recovering microbially produced IL2 in a highly pure form from the cellular material of the microorganisms that produced it comprising: disrupting the cell membranes of the microorganisms; extracting the disruptate with a chaotropic agent, such as urea, that selectively extracts microbial proteins from the cellular material; solubilizing the IL-2 in the solid phase of the extraction, mixture with an aqueous solution of a solubilizing agent, such as SDS, containing a reducing agent; and separating the IL-2 from the resulting solution by an optional extraction with 2butanol or 2-methyl-2-butanol followed by gel filtration chromatography, oxidizing the IL-2 and purifying the oxidized IL-2 by RP-HPLC.
5420009 DETECTION OF X A N T H O M O N A S CAMPESTRIS PV. CITRI BY HYBRIDIZATION AND P O L Y M E R A S E CHAIN R E A C T I O N ASSAYS Hartung John S; Pruvost Olivier Beltsville, MD, UNITED STATES Assigned to The United States of America as represented by the Secretary of Agriculture Xanthomonas campestris pv. cirri is a quarantine organism under United States and International law because of the serious disease of citrus, citrus bacterial canker disease, which is caused by the organism. We have cloned, in vector pUC9, a 4.2-
kb BamHl fragment of plasmid DNA from a typical strain of this pathogen and demonstrated that this DNA fragment specifically identifies the pathogen. The procedure involves isolation and cultivation of the bacterium, chemical isolation of its DNA, digestion of the DNA by restriction endonucleases and analysis by Southern or dot blotting using the cloned DNA fragment as biotinlabeled hybridization probe. A subclone has been made from the original 4.2-kb BamHI fragment which has sensitivity and specificity equal or greater than the original clone and which is approximately 572 bp in length. All tested strains of the most virulent form of the pathogen, type A, have a BamHI fragment of 4.2-kb which hybridizes with either probe. Other less pathogenic forms of the bacterium have BamHl fragments greater than 20kb in size. Thus not only are all strains of the pathogen detected with this probe, but sub-pathovar assignment of unknown strains is also facilitated. Strains of X. campestris which cause another non-threatening disease of citrus, citrus bacterial spot disease, are not detected by the probes. This will allow rapid, sensitive and specific detection of the pathogen in groves or from commercial shipments of citrus. In addition, oligonucleotide primers were designed, based on the nucleotide sequence of the 572-bp probe. The primers are effective in the amplification of DNA from the bacterium; thereby increasing both the specificity and sensitivity of detection methods.
5420010 ASSAY FOR E V A L U A T I N G INHIBITION OF POLYMORPHONUCLEAR L E U K O C Y T E E L A S T A S E BY NSUBSTITUTED A Z E T I D I N O N E S Finke Paul E; Hagmann William; Hanlon William; Humes John; Knight Wilson; MacCoss Malcol; Mumford Richard; Shah Shrenik K Milltosn, N J, UNITED STATES Assigned to Merck & Co Inc N-substituted azetidinones are a class of inhibitors of human leukocytes elastase which are known to be useful in the treatment of a wide variety of antiinflammatory and antidegenerative diseases. In inhibiting elastase, the therapeutic agents are shown to form a characteristic stable complex with the enzyme. In the assay disclosed herein, the inhibitor-enzyme complex is advantageously hydrolyzed and specific product(s) of the hydrolysis are measured. The assays are useful in
PATENT ABSTRACTS
working under vacuum which remove the ether after the completion of treatment; a gas chromatograph with automatic sampling for determining the residues of ether and the safety of the treated components; and mechanisms for returning the treated and safe components, in conjunction with intravenous fluids, separately or together with the healthy components, to the patient. The treatment cycle is repeated until the patient's blood is made free of virus.
5419899 PURIFIED, R E D U C E D R E C O M B I N A N T INTERLEUKIN-2 COMPOSITIONS Koths Kirston; Thomson James; Kunitani Michael; Wilson Kenneth; Hanisch Wolf Berkeley, CA, 94702, UNITED STATES A process for recovering microbially produced IL2 in a highly pure form from the cellular material of the microorganisms that produced it comprising: disrupting the cell membranes of the microorganisms; extracting the disruptate with a chaotropic agent, such as urea, that selectively extracts microbial proteins from the cellular material; solubilizing the IL-2 in the solid phase of the extraction, mixture with an aqueous solution of a solubilizing agent, such as SDS, containing a reducing agent; and separating the IL-2 from the resulting solution by an optional extraction with 2butanol or 2-methyl-2-butanol followed by gel filtration chromatography, oxidizing the IL-2 and purifying the oxidized IL-2 by RP-HPLC.
5420009 DETECTION OF X A N T H O M O N A S CAMPESTRIS PV. CITRI BY HYBRIDIZATION AND P O L Y M E R A S E CHAIN R E A C T I O N ASSAYS Hartung John S; Pruvost Olivier Beltsville, MD, UNITED STATES Assigned to The United States of America as represented by the Secretary of Agriculture Xanthomonas campestris pv. cirri is a quarantine organism under United States and International law because of the serious disease of citrus, citrus bacterial canker disease, which is caused by the organism. We have cloned, in vector pUC9, a 4.2-
kb BamHl fragment of plasmid DNA from a typical strain of this pathogen and demonstrated that this DNA fragment specifically identifies the pathogen. The procedure involves isolation and cultivation of the bacterium, chemical isolation of its DNA, digestion of the DNA by restriction endonucleases and analysis by Southern or dot blotting using the cloned DNA fragment as biotinlabeled hybridization probe. A subclone has been made from the original 4.2-kb BamHI fragment which has sensitivity and specificity equal or greater than the original clone and which is approximately 572 bp in length. All tested strains of the most virulent form of the pathogen, type A, have a BamHI fragment of 4.2-kb which hybridizes with either probe. Other less pathogenic forms of the bacterium have BamHl fragments greater than 20kb in size. Thus not only are all strains of the pathogen detected with this probe, but sub-pathovar assignment of unknown strains is also facilitated. Strains of X. campestris which cause another non-threatening disease of citrus, citrus bacterial spot disease, are not detected by the probes. This will allow rapid, sensitive and specific detection of the pathogen in groves or from commercial shipments of citrus. In addition, oligonucleotide primers were designed, based on the nucleotide sequence of the 572-bp probe. The primers are effective in the amplification of DNA from the bacterium; thereby increasing both the specificity and sensitivity of detection methods.
5420010 ASSAY FOR E V A L U A T I N G INHIBITION OF POLYMORPHONUCLEAR L E U K O C Y T E E L A S T A S E BY NSUBSTITUTED A Z E T I D I N O N E S Finke Paul E; Hagmann William; Hanlon William; Humes John; Knight Wilson; MacCoss Malcol; Mumford Richard; Shah Shrenik K Milltosn, N J, UNITED STATES Assigned to Merck & Co Inc N-substituted azetidinones are a class of inhibitors of human leukocytes elastase which are known to be useful in the treatment of a wide variety of antiinflammatory and antidegenerative diseases. In inhibiting elastase, the therapeutic agents are shown to form a characteristic stable complex with the enzyme. In the assay disclosed herein, the inhibitor-enzyme complex is advantageously hydrolyzed and specific product(s) of the hydrolysis are measured. The assays are useful in