Abstract: Eucaryotic cells cotransformed with product and selection genes yield considerably greater quantities of product after a novel subcloning strategy is employed: Transformants are identified for product yield, cultured under selection pressure and the progeny screened for product yield. Novel transformation vectors contain directly ligated selection and product genes and/or eucaryotic promoters.

Abstract: RNA such as messenger RNA is digested to nucleotide phosphates including AMP or ADP. The ATP or a byproduct of the phosphorylation, e.g., pyruvate, is detected. Exemplary enzymes used (with appropriate co-reactants and co-factors) are: (1) polynucleotide phosphorylase, pyruvate kinase and luciferase, or (2) phosphodiesterase (or RNase), myokinase, pyruvate kinase and luciferase. The phosphorylation to ATP (e.g., with pyruvate kinase) is preferably coupled with the previous (reversible) enzymatic step.

Abstract: Reagent complexes containing a probe polynucleotide with at least two target binding regions. A labeled polynucleotide is bound to at least a portion of each target binding region. In one method, sample nucleic acid displaces labeled polynucleotide from one target binding region; after washing, a reagent polynucleotide displaces labeled polynucleotide from the other target binding region. In a second method, sample nucleic acid strands having two target nucleic acid sequences in proximity (e.g., sequences translocated in certain cancers) displace the labeled polynucleotide from both target binding region.

Abstract: A target nucleotide sequence having a half-restriction site is determined in the nucleic acid of a biological sample. It is contacted with a reagent complex of:(i) a labeled probe polynucleotide containing a target binding region which is substantially complementary to the target nucleotide sequence and which contains a unique half-restriction site completely complementary to the half-restriction site of the target nucleotide sequence; and(ii) a second polynucleotide hybridized to the labeled probe polynucleotide in at least a portion of the target binding region, the portion including the unique half-restriction site of the target binding region;The second polynucleotide contains at least one mismatched or unpaired nucleotide opposite to the unique half-restriction site of the labeled probe polynucleotide, whereby a restriction enzyme specific for the unique restriction site will not cleave the reagent complex.

Abstract: A method for purifying erythropoietin is described. The method comprises treating partially purifying erythropoietin by reverse phase high performance liquid chromatography to obtain homogeneous erythropoietin having a molecular weight of about 34,000 daltons on SDS PAGE and moving a single peak on reverse phase HPLC. The homogeneous erythropoietin protein preferably has a specific activity of at least 120,000 IU, more preferably at least 160,000 IU per absorbance unit at 280 nm.

Abstract: A method for identifying and isolating clones containing DNA coding for a desired protein is described. DNA prepared from a cell that expresses the desired protein is inserted into an isolation expression vector having means for replication (as a means of producing DNA) and a suitable promoter for expression of said DNA in a predetermined mammalian host cell as well as means for replication in a bacterial cell. The transient expression vector is then inserted into a bacterial cell for replication of the DNA. Pools of DNA, prepared from a predetermined number of bacterial clones so that the nucleic acids (DNA and RNA) is substantially free of other bacterial contaminants are transfected or microinjected into mammalian host cells and conditioned medium from growing such cells is tested for the presence of the desired protein. Positive pools are selected and the clones used to make the pool are screened to identify and isolate the clone containing the desired DNA.

Abstract: A support system for organic synthesis comprising magnetic particles in a dispersion medium covalently attached to functional groups having affinity for polymer subunits, and methods for making and using the support system, e.g. for synthesis of oligodeoxynucleotides and polypeptides.

Abstract: A process is described for producing L-4-phenyl-2-amino butanoic acid. The process comprises reacting 4-phenyl-2-oxobutanoic acid or an ester thereof with L-aspartic acid in the presence of transaminase enzyme to produce L-4-phenyl-2-aminobutanoic acid and oxaloacetate, and decarboxylating said oxaloacetate.

Abstract: A process is described for producing alpha amino acids or derivatives thereof. The process comprises reacting an alpha-keto acid with L-aspartic acid in the presence of transaminase enzyme to produce (1) an alpha amino acid corresponding to said alpha-keto acid and (2) oxaloacetate; and decarboxylating said oxaloacetate.