Abstract: Novel recombinant DNA cosmid shuttle vectors and a method of using them in the construction of genomic DNA libraries are described. The vectors demonstrate the incorporation of both the size selection and in vitro packaging mechanisms of lambda into a Streptomyces-E. coli shuttle vector by the incorporation of two or more COS sequences of bacteriophage lambda.

Abstract: Recombinant DNA plasmids capable of functioning as shuttle vectors in Rhodococcus equi, Corynebacterium, E. coli, B. subtilis, or S. aureus are disclosed. The recombinant plasmid contains an origin of replication which is functional in Rhodococcus and at least one of E. coli, Corynebacterium, S. aureus, and B. subtilis. Additionally, the plasmid contains a heritable, selectable marker.

Abstract: DNA sequences encoding a protein having angiogenic activity are disclosed. Expression vectors containing these sequences are introduced into host cells and direct the production of the angiogenic protein. Proteins produced according to the invention are useful in the diagnosis of malignancies, for promoting wound healing, and for other diagnostic and therapeutic purposes.

Abstract: A class of subtilisin analogs suitable for admixture to cleaning compositions and having improved stability over naturally occurring Bacillus subtilisins are prepared by expressing a modified gene encoding the subtilisin analog in Bacillus subtilis. The subtilisin analogs are characterized as having a modified calcium binding site to improve calcium binding and either an Asn or a Gly replaced in any Asn-Gly sequences present in the subtilisin.

Abstract: An aqueous suspension of micro-organism cells containing a 3-hydroxybutyrate polymer are subjected to a proteolytic enzyme digestion and/or a surfactant digestion in order to solubilise cell material other than the 3-hydroxybutyrate polymer.Prior to, or during the digestion, but before any proteolytic enzyme digestion step, the suspension is heated to at least 80.degree. C. to denature nucleic acids which otherwise hinder separation of the 3-hydroxybutyrate polymer containing residue from the suspension.

Abstract: A mixture of Pseudomonas putida one having plasmids encoding the camphor (CAM) and toluene (TOL) degradation and the other having a plasmid encoding for naphthalene (NAH) degradation is described. The mixture is more effective than either Pseudomonas putida alone or than a single Pseudomonas putida with three related plasmids which encode for the degradation of CAM, TOL and NAH.

Abstract: The invention concerns a method for extending the half-life of mRNAs. The half-life extension is conferred upon the mRNA by a co-transcribed positive retroregulatory element which is ligated to the 3' end of the DNA sequence encoding the RNA. RNAs having an extended half-life conferred by a co-transcribed positive retroregulatory element are also claimed.

Abstract: A method of continuous product formation using at least two continuous fermentation units and a microorganism capable of being induced, in response to environmental conditions, to undergo a genetic alteration from a state favoring microorganism growth to a state favoring product production by the microorganism. The first continuous fermentation unit is maintained at environmental conditions selected to favor growth of the microorganism and to be nonpermissive for the genetic alteration. The microorganism is grown continuously in the first unit, and a portion of the growing microorganism cell mass is transferred via connecting means to the second continuous fermentation unit. Either the connecting means or the second unit is maintained at second environmental conditions selected to effect the genetic alteration. The altered microorganism is cultured in the second unit.

Abstract: This discovery resulted when a .lambda. gt11 cDNA library of normal human melanocytes were screened with antibodies directed against purified hamster tyrosinase. Sixteen independent clones which gave a positive signal were isolated from 5.times.10.sup.5 independent plaques. cDNA inserts of 13 clones among the 16 candidates cross-hybridized with each other, indicating that they were from related mRNA species. mRNA homologous to a representative cDNA .lambda. mel 34 was expressed specifically in melanocytes, detecting an approximately 2.4 kb mRNA species of human melanocytes. The nucleotide sequence of the three overlapping cDNA inserts spanning 1.88 kb was determined and an amino acid sequence was deducted. The human tyrosinase is composed of 548 amino acids with a molecular weight of 62,160 excluding a hydrophobic signal peptide. Mouse genomic DNA blot analysis revealed that the gene for .lambda. mel 34 was deleted in albino mouse homozygous for the deletion at and around the albino locus on chromosome 7.

Abstract: A process is provided for producing an ordered series of cloned, circular, DNA molecules containing shortened target DNA segments derived from a long target DNA segment, which are suitable for use in determining the nucleotide sequence of the long target DNA segment, or for targeting specific regions within the target DNA segment. The process includes producing, by molecular cloning, a plurality of double-stranded recombinant DNA molecules each containing: (i) vector DNA; (ii) a sequencing primer binding site; and, (iii) a DNA region having unique endonuclease sites and a long target DNA segment. The sequencing primer binding site is spaced from the long target DNA segment by at least a portion of said DNA region having the unique endonuclease sites. The plurality of double-stranded circular recombinant DNA molecules are cleared using two restriction endonucleases.

Abstract: Disclosed is a heterodimeric T lymphocytes receptor subunit. The subunit consists of variable, joining, constant, transmembrane, and cytoplasmic regions.The structure, amino acid, and nucleotide sequence of the lymphocyte receptor subunit were determined using cDNA clones derived from a functional murine cytotoxic T lymphocyte clone. The genes corresponding to these cDNA are expressed and rearranged specifically in T cells and have significant sequence homologies to immunoglobulin V and C genes.T cell receptor subunits may be produced from the cDNA clones. The protein molecules may be further used for the production of T-cell clone specific antibodies.

Abstract: Disclosed is a heterodimeric T lymphocyte receptor comprising an alpha and a beta subunit. Each subunit consists of a signal peptide, variable, joining, constant, transmembrane, and cytoplasmic regions. The two subunits are connected by a disulfide bond between cysteine residues located between the constant and transmembrane region.The structure, amino acid, and nucleotide sequence of the lymphocyte receptor were determined using cDNA clones derived from a functional murine cytotoxic T lymphocyte clone. The genes corresponding to these cDNA are expressed and rearranged specifically in T cells and have significant sequence homologies to immunoglobulin V and C genes.Both the T cell receptor protein and its subunits may be produced from the cDNA clones. The protein molecules may be further used for the production of T-cell clone specific antibodies.

Abstract: The invention concerns a double-stranded recombinant DNA cloning vector comprising one single insertion site for foreign DNA and one or two labelling sites adjacent or approximate to said insertion site. In the instance of two labelling sites, said sites flank the insertion site. The labelling site(s) can be individually labelled in a fashion that results in the labelling of the 3' end of either strand. The use of the invention in a process affords the ability to individually label only one strand at the 3' end in a simplified method for base sequencing analysis.

Abstract: The present invention provides a micro-organism of the species Escherichia coli or Pseudomonas putida, wherein it constitutively forms creatinamidinohydrolase.The present invention also provides a process for the production of such a micro-organism.

Abstract: A process is disclosed for purifying a recombinant fusion protein having an N-terminal identification sequence comprising multiple anionic amino acid residues, comprising forming a complex of the protein with a divalent cation dependent monoclonal antibody specific for the sequence, isolating the complex, and dissociating antibody and protein by selectively depleting the concentration of divalent cations in contact with the complex. A particular calcium-dependent monoclonal antibody, 4E11, is useful in an embodiment of the process which employs the identification peptide DYKDDDDK.

Abstract: A novel method of selecting Streptomyces recombinant DNA-containing host cells and vectors useful in exemplifying the method are described. The vectors confer apramycin resistance to sensitive Streptomyces host cells and thus provide a convenient method of selecting Streptomyces transformants. The apramycin resistance-conferring gene used in the method is an acetyltransferase aac(3)IV gene and can be isolated from E. coli K12 BE1041/pKC309 (NRRL B-15827) on an .about.1.5 kb PstI-EcoRI restriction fragment.

Abstract: A process is provided for producing an ordered series of cloned, circular, DNA molecules containing shortened target DNA segments derived from a long target DNA segment, which are suitable for use in determining the nucleotide sequence of the long target DNA segment, or for targeting specific regions within the target DNA segment. The process includes producing, by molecular cloning, a plurality of double-stranded recombinant DNA molecules each containing: (i) vector DNA; (ii) a sequencing primer binding site; and, (iii) a DNA region having unique endonuclease sites and a long target DNA segment. The sequencing primer binding site is spaced from the long target DNA segment by at least a portion of said DNA region having the unique endonuclease sites. The plurality of double-stranded circular recombinant DNA molecules are cleared using two restriction endonucleases.

Abstract: DNA which encodes the polypeptide streptavidin has been isolated as a fragment 2 kb in length derived from a restriction endonuclease digestion of the chromosomal DNA of Streptomyces avidinii. The nucleic acid sequence of the gene and the amino acid sequence of the polypeptide have been determined. A fused gene has been prepared which comprises the streptavidin gene fused to a gene encoding the human LDL receptor. Expression of the gene fusion results in a fused streptavidin-human LDL receptor polypeptide. Methods are provided for using the fused gene to produce labeled, chemically modified proteins in vivo and to isolate a protein knowing only the nucleotide sequence of the gene encoding the protein.

Abstract: A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C.sub.1 -utilizing host and in a C.sub.1 -utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C.sub.1 -utilizing host to the C.sub.1 -utilizing host; DNA providing resistance to two antibiotics to which the wild-type C.sub.1 -utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C.sub.1 -utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C.sub.1 -utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C.sub.1 -utilizing (e.g., E. coli) host, and then conjugated with a selected C.sub.1 -utilizing mutant.

Abstract: Hydroquinone is isolated from the reaction medium resulting from contacting cultured Mycobacterium cells containing an oxygenase with benzene and/or phenol in the presence of molecular oxygen or air and aqueous medium.