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5624829 Transformed industrial bacillus strains and methods for making and using them
682
PATENT ABSTRACTS
It has been discovered that any RNA can be targeted for cleavage by RNAase P from eukaryotic ceils, for example, human RNAase P, using a suitably designed oligoribonucleotide (external guide sequence , or EGS) to form a hybrid with the target RNA, thereby creating a substrate for cleavage by RNAase P in vitro. The EGS hydrogen bonds to the targeted RNA to form a partial tRNA like structure including the aminoacyl acceptor stem, the T stern and loop, and part of the D stem. The most efficient EGS with human RNAase P is the EGS in which the anticodon stem and loop was deleted. Modifications can also be made within the T-loop. Methods are also disclosed to randomly select and to express a suitable EGS in vivo to make a selected RNA a target for cleavage by the host cell RNAase P, thus preventing expression of the function of the target RNA. The methods and compositions should be useful to prevent the expression of disease-causing genes in vivo.
5624826 C L O N I N G V E C T O R PLASMID VECTOR-PRIMER DERIVED T H E R E F R O M AND P R E P A R A T I O N M E T H O D OF THE SAME Kato Seishi; Aoki Takashi; Umezawa Yuri Sagamihara, JAPAN Assigned to Sagami Chemical Research Center Disclosed is a cloning vector plasmid suitable for preparing cDNA banks. The cloning vector plasmid of the present invention comprises, in the order mentioned from upstream to downstream region unless otherwise specified, a promoter PR 1 acting in mammalian cells; an unique restriction enzyme site RE1 rarely existing in eukaryotic DNA and a promoter PR2 of RNA polymerase; an unique restriction enzyme site RE2 generating T-protruding dG tail; an unique arbitrary restriction enzyme site RE3; an unique arbitrary restriction enzyme site RE4 generating 3'-protruding terminus; an unique restriction enzyme site RE5 rarely existing in eukaryotic DNA and a promoter PR3 of RNA polymerase; and at least one kind of arbitrary restriction enzyme site RE6 generating 3'-protruding terminus existing at the arbitrary position before the restriction site REI; and further, at arbitrary
positions other than the above-described restriction enzyme sites and promoters, an origin OR1 for replication in mammalian cells, an origin OR2 for replication of a single-stranded phage, an origin OR3 for replication in E. coli, and a selective marker.
5624827 DNA SEQUENCES E N C O D I N G THE PLANT TOXIN GELONIN Rosenblum Michael G; Beattie Kenneth Houston, TX, UNITED STATES Assigned to Research Development Foundation The present invention provides the nucleotide sequence for a synthetic gene for the plant toxin gelonin and a process for producing, cloning and expressing this synthetic gene. The DNA sequence for a synthetic gene for gelonin as shown in sequence ID NO. 1. The present invention also provides expression vectors containing the DNA sequences for gelonin and cells transformed with these vectors. In addition, an immunotoxin comprising an antibody of conjugated to the protein gelonin.
5624829 T R A N S F O R M E D INDUSTRIAL BACILLUS STRAINS A N D M E T H O D S FOR M A K I N G A N D USING T H E M Sanders Johan P M; van den Berg Johannes A; Andreoli Peter M; Vos Yvonne; van Ee Jan; Mulleners Leo J S Delft, NETHERLANDS Assigned to Gist-Brocades B V Novel methods and novel industrial unicellular microorganism strains, particularly industrial Bacillus strains, are provided for enhanced production of endogenous and exogenous polypeptides. Cloning vehicles containing the gene expressing the polypeptide of interest are introduced into a compatible host. Transformed hosts harboring the introduced vehicle in a stable way by integration of the vehicle into the host cells chromosome are selected. Efficient transfer of the vehicle containing the gene of interest is achieved with the resulting industrial strain
PATENT ABSTRACTS
transformants being effective, stable producers of the desired polypeptide product.
5624830 CYTOSINE
AND THERAPIES
Mullen Craig A; Blaese R Michael Bethesda, MD, UNITED STATES Assigned to The United States of America as represented by the Department of Health and Human Services The present invention relates to a system comprising a modified bacterial gene for cytosine deaminase that has been engineered into a eukaryotic expression vector and the expression of the gene by mammalian cells. The present invention further relates to methods, gene therapies and vaccines that employ the negative selectable marker, cytosine deaminase, which has the ability to produce a toxic antimetabolic 5-fluorouracil from 5-fluorocytosine.
5624832 BETA1
5624833 PURIFIED THERMOSTABLE NUCLEIC ACID POLYMERASE ENZYME THERMOTOGA
FROM MARITIMA
Gelfand David H; Lawyer Frances; Stoffel Susanne Oakland, CA, UNITED STATES Assigned to Hoffmann-La Roche Inc A purified thermostable enzyme is derived from the eubacterium Thermotoga maritima. The enzyme has a molecular weight as determined by gel electrophoresis of about 97 kilodaltons and DNA polymerase I activity. The enzyme can be produced from native or recombinant host cells and can be used with primers and nucleoside triphosphates in a temperature-cycling chain reaction where at least one nucleic acid sequence is amplified in quantity from an existing sequence.
6
N-ACETYLGLUCOSAMINYLTR ANSFERASE, ITS ACCEPTOR MOLECULE, LEUKOSIALIN, AND A METHOD FOR CLONING PROTEINS HAVING ENZYMATIC
using CHO cells that support replication of plasmid vectors having a polyoma virus origin of replication. A method to obtain a suitable cell line that expresses an acceptor molecule also is disclosed.
DEAMINASE
NEGATIVE SELECTION SYSTEM FOR GENE TRANSFER TECHNIQUES
683
ACTIVITY
Fukuda Minom; Bierhuizen Marti F San Diego, CA, UNITED STATES Assigned to La Jolla Cancer Research Foundation The present invention provides a novel betal right arrow6 N-acetylglucosaminyltransferase, which forms core 2 oligosaccharide structures in O-glycans, and a novel acceptor molecule, leukosialin, CD43, for core 2 betal right arrow6 N-acetylglucosaminyltransferase activity. The amino acid sequences and nucleic acid sequences encoding these molecules, as well as active fragments thereof, also are disclosed. A method for isolating nucleic acid sequences encoding proteins having enzymatic activity is disclosed,
5624835 ENDO-BETA-1,4-GLUCANASE AND A DNASEQUENCE Duml orreich Kur; Christensen Flemming M; Schnell Yvette; Mischler Marcel; Dalbge Henrik; Heldt-Hansen Hans Pns Ps P Grenzach Wyhlen, GERMANY Assigned to Novo Nordisk A/S PCT No. PCT/DK93/00108 Sec. 371 Date Oct. 5, 1994 Sec. 102(e) Date Oct. 5, 1994 PCT Filed Mar. 26, 1993 PCT Pub. No. WO93/20193 PCT Pub. Date Oct. 14, 1993. The molecular characteristics and a partial amino acid sequence of an endo- beta-l,4-glucanase obtainable from Aspergillus aculeatus are described, as well as corresponding recombinant DNA sequences, vectors, and transformed hosts. Use of the endobeta-l,4-glucanase or a pectinase preparation enriched with the endo- beta-l,4-glucanase for degradation or modification of plant cell walls is described.
PATENT ABSTRACTS
It has been discovered that any RNA can be targeted for cleavage by RNAase P from eukaryotic ceils, for example, human RNAase P, using a suitably designed oligoribonucleotide (external guide sequence , or EGS) to form a hybrid with the target RNA, thereby creating a substrate for cleavage by RNAase P in vitro. The EGS hydrogen bonds to the targeted RNA to form a partial tRNA like structure including the aminoacyl acceptor stem, the T stern and loop, and part of the D stem. The most efficient EGS with human RNAase P is the EGS in which the anticodon stem and loop was deleted. Modifications can also be made within the T-loop. Methods are also disclosed to randomly select and to express a suitable EGS in vivo to make a selected RNA a target for cleavage by the host cell RNAase P, thus preventing expression of the function of the target RNA. The methods and compositions should be useful to prevent the expression of disease-causing genes in vivo.
5624826 C L O N I N G V E C T O R PLASMID VECTOR-PRIMER DERIVED T H E R E F R O M AND P R E P A R A T I O N M E T H O D OF THE SAME Kato Seishi; Aoki Takashi; Umezawa Yuri Sagamihara, JAPAN Assigned to Sagami Chemical Research Center Disclosed is a cloning vector plasmid suitable for preparing cDNA banks. The cloning vector plasmid of the present invention comprises, in the order mentioned from upstream to downstream region unless otherwise specified, a promoter PR 1 acting in mammalian cells; an unique restriction enzyme site RE1 rarely existing in eukaryotic DNA and a promoter PR2 of RNA polymerase; an unique restriction enzyme site RE2 generating T-protruding dG tail; an unique arbitrary restriction enzyme site RE3; an unique arbitrary restriction enzyme site RE4 generating 3'-protruding terminus; an unique restriction enzyme site RE5 rarely existing in eukaryotic DNA and a promoter PR3 of RNA polymerase; and at least one kind of arbitrary restriction enzyme site RE6 generating 3'-protruding terminus existing at the arbitrary position before the restriction site REI; and further, at arbitrary
positions other than the above-described restriction enzyme sites and promoters, an origin OR1 for replication in mammalian cells, an origin OR2 for replication of a single-stranded phage, an origin OR3 for replication in E. coli, and a selective marker.
5624827 DNA SEQUENCES E N C O D I N G THE PLANT TOXIN GELONIN Rosenblum Michael G; Beattie Kenneth Houston, TX, UNITED STATES Assigned to Research Development Foundation The present invention provides the nucleotide sequence for a synthetic gene for the plant toxin gelonin and a process for producing, cloning and expressing this synthetic gene. The DNA sequence for a synthetic gene for gelonin as shown in sequence ID NO. 1. The present invention also provides expression vectors containing the DNA sequences for gelonin and cells transformed with these vectors. In addition, an immunotoxin comprising an antibody of conjugated to the protein gelonin.
5624829 T R A N S F O R M E D INDUSTRIAL BACILLUS STRAINS A N D M E T H O D S FOR M A K I N G A N D USING T H E M Sanders Johan P M; van den Berg Johannes A; Andreoli Peter M; Vos Yvonne; van Ee Jan; Mulleners Leo J S Delft, NETHERLANDS Assigned to Gist-Brocades B V Novel methods and novel industrial unicellular microorganism strains, particularly industrial Bacillus strains, are provided for enhanced production of endogenous and exogenous polypeptides. Cloning vehicles containing the gene expressing the polypeptide of interest are introduced into a compatible host. Transformed hosts harboring the introduced vehicle in a stable way by integration of the vehicle into the host cells chromosome are selected. Efficient transfer of the vehicle containing the gene of interest is achieved with the resulting industrial strain
PATENT ABSTRACTS
transformants being effective, stable producers of the desired polypeptide product.
5624830 CYTOSINE
AND THERAPIES
Mullen Craig A; Blaese R Michael Bethesda, MD, UNITED STATES Assigned to The United States of America as represented by the Department of Health and Human Services The present invention relates to a system comprising a modified bacterial gene for cytosine deaminase that has been engineered into a eukaryotic expression vector and the expression of the gene by mammalian cells. The present invention further relates to methods, gene therapies and vaccines that employ the negative selectable marker, cytosine deaminase, which has the ability to produce a toxic antimetabolic 5-fluorouracil from 5-fluorocytosine.
5624832 BETA1
5624833 PURIFIED THERMOSTABLE NUCLEIC ACID POLYMERASE ENZYME THERMOTOGA
FROM MARITIMA
Gelfand David H; Lawyer Frances; Stoffel Susanne Oakland, CA, UNITED STATES Assigned to Hoffmann-La Roche Inc A purified thermostable enzyme is derived from the eubacterium Thermotoga maritima. The enzyme has a molecular weight as determined by gel electrophoresis of about 97 kilodaltons and DNA polymerase I activity. The enzyme can be produced from native or recombinant host cells and can be used with primers and nucleoside triphosphates in a temperature-cycling chain reaction where at least one nucleic acid sequence is amplified in quantity from an existing sequence.
6
N-ACETYLGLUCOSAMINYLTR ANSFERASE, ITS ACCEPTOR MOLECULE, LEUKOSIALIN, AND A METHOD FOR CLONING PROTEINS HAVING ENZYMATIC
using CHO cells that support replication of plasmid vectors having a polyoma virus origin of replication. A method to obtain a suitable cell line that expresses an acceptor molecule also is disclosed.
DEAMINASE
NEGATIVE SELECTION SYSTEM FOR GENE TRANSFER TECHNIQUES
683
ACTIVITY
Fukuda Minom; Bierhuizen Marti F San Diego, CA, UNITED STATES Assigned to La Jolla Cancer Research Foundation The present invention provides a novel betal right arrow6 N-acetylglucosaminyltransferase, which forms core 2 oligosaccharide structures in O-glycans, and a novel acceptor molecule, leukosialin, CD43, for core 2 betal right arrow6 N-acetylglucosaminyltransferase activity. The amino acid sequences and nucleic acid sequences encoding these molecules, as well as active fragments thereof, also are disclosed. A method for isolating nucleic acid sequences encoding proteins having enzymatic activity is disclosed,
5624835 ENDO-BETA-1,4-GLUCANASE AND A DNASEQUENCE Duml orreich Kur; Christensen Flemming M; Schnell Yvette; Mischler Marcel; Dalbge Henrik; Heldt-Hansen Hans Pns Ps P Grenzach Wyhlen, GERMANY Assigned to Novo Nordisk A/S PCT No. PCT/DK93/00108 Sec. 371 Date Oct. 5, 1994 Sec. 102(e) Date Oct. 5, 1994 PCT Filed Mar. 26, 1993 PCT Pub. No. WO93/20193 PCT Pub. Date Oct. 14, 1993. The molecular characteristics and a partial amino acid sequence of an endo- beta-l,4-glucanase obtainable from Aspergillus aculeatus are described, as well as corresponding recombinant DNA sequences, vectors, and transformed hosts. Use of the endobeta-l,4-glucanase or a pectinase preparation enriched with the endo- beta-l,4-glucanase for degradation or modification of plant cell walls is described.