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4710465 Junction-fragment DNA probes and probe clusters
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PATENT ABSTRACTS 4710465
4710471
JUNCTION-FRAGMENT DNA PROBES AND PROBE CLUSTERS
NOVEL VECTOR PLASMIDS
Sherman M Weissman, FranciCollins assigned to Yale University A junction-fragment DNA probe, a DNA probe cluster, and methods of preparing and using the probe and cluster to study gene localization and organization. The probe includes first and second gene segments which are derived from first and second, single-copy genomic-DNA gene regions, respectively, separated from one another, in the genomic DNA strand, by a selected distance of between about 20 and 2,000 kilobases. The two segments in the probe are connected at a junction region which may include a marker segment usable in isolating and/or selecting the probe. The probe cluster includes a set of such probes, each having a common first segment, and a second segment which is derived from one of a number of gene regions located at various distances from the first gene region in the genomic DNA strand from which the two segments are derived.
Ryoichi Katsumata, Akio Ozaki, Tetsuo Oka, Akira Furuya, Mashida, Japan assigned to Kyowa Hakko Kogyo Co Ltd Disclosed are recombinant plasmid vectors constructed of a plasmid autonomously replicable in cells of microorganisms belonging to the genus Corynebacterium or Brevibacterium and a DNA fragment containing a gene expressible by said microorganisms. The recombinant vector plasmids are autonomously replicable in glutamic acid-producing microorganisms and are useful to clone desired DNA fragments in such microorganisms.
4710473 DNA PLASMIDS Charles Morris assigned to Amgen Inc
4710466 METHOD OF CLONING MODIFIED STREPTOMYCETES DNA Charles L Hershberger, Jeffrey L Larson assigned to Eli Lilly and Company A method of cloning endogenously modified Streptomycetes DNA, which is normally rejected by restrictionless heterospecific hosts, is disclosed. The method uses bacteriophage lambda to construct a genomic library of modified Streptomycetes DNA; such lambda-containing Streptomycetes DNA is replicated to provide a source of non-modified Streptomycetes DNA. This non-modified DNA is subcloned into a selectable cloning vector and used to transform restrictionless hetero-specific hosts. The transformants can then be screened for clones containing genes of interest.
Disclosed are novel circular DNA plasmids useful as vectors in recombinant methods to secure high levels of E.coli expression of exogenous genes. Plasmids of the invention comprise discrete DNA sequences operative to: (1) confer upon the plasmid the capacity for autonomous replication in a host cell; (2) control autonomous plasmid replication in relation to the temperature at which host cell cultures are maintained; (3) stabilize maintenance of the plasmid in host cell populations; (4) direct synthesis of a protein product indicative of plasmid maintenance in a host cell population; (5) provide, in series, a plurality of restriction endonuclease recognition sites, unique to the plasmid and facilitative of exogenous gene DNA sequence insertion; and (6) terminate mRNA transcription of adjacent DNA sequences and situated so as to terminate transcription of exogenous gene sequences inserted within the plasmid at said unique restriction endonuclease restriction sites. Plasmids preferably have a size of less than 5.0 kilobases (exclusive of any inserted exogenous gene) and optionally include a DNA sequence operative to provide a strong promoter of mRNA transcription functionally associated with a temperature sensitive repressor sequence. A presently preferred embodiment of novel plasmids of the invention is plasmid pCFM414 (A.T.C.C. No. 40076).
PATENT ABSTRACTS 4710465
4710471
JUNCTION-FRAGMENT DNA PROBES AND PROBE CLUSTERS
NOVEL VECTOR PLASMIDS
Sherman M Weissman, FranciCollins assigned to Yale University A junction-fragment DNA probe, a DNA probe cluster, and methods of preparing and using the probe and cluster to study gene localization and organization. The probe includes first and second gene segments which are derived from first and second, single-copy genomic-DNA gene regions, respectively, separated from one another, in the genomic DNA strand, by a selected distance of between about 20 and 2,000 kilobases. The two segments in the probe are connected at a junction region which may include a marker segment usable in isolating and/or selecting the probe. The probe cluster includes a set of such probes, each having a common first segment, and a second segment which is derived from one of a number of gene regions located at various distances from the first gene region in the genomic DNA strand from which the two segments are derived.
Ryoichi Katsumata, Akio Ozaki, Tetsuo Oka, Akira Furuya, Mashida, Japan assigned to Kyowa Hakko Kogyo Co Ltd Disclosed are recombinant plasmid vectors constructed of a plasmid autonomously replicable in cells of microorganisms belonging to the genus Corynebacterium or Brevibacterium and a DNA fragment containing a gene expressible by said microorganisms. The recombinant vector plasmids are autonomously replicable in glutamic acid-producing microorganisms and are useful to clone desired DNA fragments in such microorganisms.
4710473 DNA PLASMIDS Charles Morris assigned to Amgen Inc
4710466 METHOD OF CLONING MODIFIED STREPTOMYCETES DNA Charles L Hershberger, Jeffrey L Larson assigned to Eli Lilly and Company A method of cloning endogenously modified Streptomycetes DNA, which is normally rejected by restrictionless heterospecific hosts, is disclosed. The method uses bacteriophage lambda to construct a genomic library of modified Streptomycetes DNA; such lambda-containing Streptomycetes DNA is replicated to provide a source of non-modified Streptomycetes DNA. This non-modified DNA is subcloned into a selectable cloning vector and used to transform restrictionless hetero-specific hosts. The transformants can then be screened for clones containing genes of interest.
Disclosed are novel circular DNA plasmids useful as vectors in recombinant methods to secure high levels of E.coli expression of exogenous genes. Plasmids of the invention comprise discrete DNA sequences operative to: (1) confer upon the plasmid the capacity for autonomous replication in a host cell; (2) control autonomous plasmid replication in relation to the temperature at which host cell cultures are maintained; (3) stabilize maintenance of the plasmid in host cell populations; (4) direct synthesis of a protein product indicative of plasmid maintenance in a host cell population; (5) provide, in series, a plurality of restriction endonuclease recognition sites, unique to the plasmid and facilitative of exogenous gene DNA sequence insertion; and (6) terminate mRNA transcription of adjacent DNA sequences and situated so as to terminate transcription of exogenous gene sequences inserted within the plasmid at said unique restriction endonuclease restriction sites. Plasmids preferably have a size of less than 5.0 kilobases (exclusive of any inserted exogenous gene) and optionally include a DNA sequence operative to provide a strong promoter of mRNA transcription functionally associated with a temperature sensitive repressor sequence. A presently preferred embodiment of novel plasmids of the invention is plasmid pCFM414 (A.T.C.C. No. 40076).