Abstract: Illustrated is the preparation and expression of manufactured genes capable of directing synthesis of human immune and leukocyte interferons and of other biologically active proteinaceous products, which products differ from naturally-occurring forms in terms of the identity and/or relative position of one or more amino acids, and in terms of one or more biological and pharmacological properties but which substantially retain other such properties.

Abstract: Illustrated is the preparation and expression of manufactured genes capable of directing synthesis of human immune and leukocyte interferons and of other biologically active proteinaceous products, which products differ from naturally-occurring forms in terms of the identity and/or relative position of one or more amino acids, and in terms of one or more biological and pharmacological properties but which substantially retain other such properties.

Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences in excess of about 200 base pairs in length, which sequences may comprise entire structural genes. Novel sequences are prepared from two or more DNA subunits provided with terminal regions comprising restriction endonuclease cleavage sites facilitating insertion of subunits into a selected vector for purposes of amplification during the course of the total assembly process. The total, finally-assembled sequences include at least one, and preferably two or more, unique restriction endonuclease cleavage site(s) at intermediate positions along the sequence, allowing for easy excision and replacement of subunits and the correspondingly facile preparation of multiple structural analogs of polypeptides coded for by the sequences.

Abstract: Disclosed are analogs of human leukocyte interferon which predominantly include those amino acid residues which are common to all naturally-occurring human leukocyte interferon subtype amino acid sequences and which include, at one or more of those positions where there is no amino acid common to all subtypes, an amino acid which predominantly occurs at that position and in no event include any amino acid residue which is not extant in that position in at least one naturally-occurring subtype.

Abstract: A mutated subtilisin suitable for admixture to washing compositions and exhibiting substantially improved stability over naturally occurring Bacillus serine proteases is prepared by expressing a modified gene encoding subtilisin in Bacillus subtilis. A preferred subtilisin analog product differs from wild-type Bacillus alkaline proteases by having any amino acid, and preferably serine, at position 218 in place of asparagine. The product is preferably produced in a strain of B. subtilis which is mutated to block synthesis of endogenous proteases. The method of replacing an Asn or a Gly in an Asn-Gly sequence in order to improve pH and thermal stability may be applied to other sites in subtilisin and to other proteins as well.

Abstract: This disclosure relates to a novel class of serine proteases isolated from a culture medium of fungus Tritirachium album. The serine proteases disclosed have a high degree of stability in detergent formulations.In addition, this disclosure relates to a process for producing such serine proteases using recombinant techniques.

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: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences in excess of about 200 base pairs in length, which sequences may comprise entire structural genes. Novel sequences are prepared from two or more DNA subunits provided with terminal regions comprising restriction endonuclease cleavage sites facilitating insertion of subunits into a selected vector for purposes of amplification during the course of the total assembly process. The total, finally-assembled sequences include at least one, and preferably two or more, unique restriction endonuclease cleavage site(s) at intermediate positions along the sequence, allowing for easy excision and replacement of subunits and the correspondingly facile preparation of multiple structural analogs of polypeptides coded for by the sequences.

Abstract: A probe is attached to a stock support to which is bound a single-stranded polynucleotide by hybridizing both the probe and the bound polynucleotide to a connecting strand with which they are both complementary and then ligating an end of the bound polynucleotide and the probe. After denaturation, the probe remains attached to the support by way of the bound polynucleotide. A target polynucleotide is then hybridized with the support-bound probe and with a labelled probe to provide a detectable hybridization complex.

Abstract: A method for the isolation and quantitative detection of a selected single-stranded target polynucleotide from solution. The target polynucleotide is hybridized in solution to a single-stranded mediator polynucleotide, a probe polynucleotide, and an immobilized polynucleotide sequence. The sequence of the mediator polynucleotide comprises a first sequence complementary to a first portion of the target polynucleotide sequence and a second nucleotide sequence complementary to a portion of a single-stranded immobilized polynucleotide sequence. The probe polynucleotide, which carries a detectable label, is complementary to a second portion of the necleotide sequence of the target. The immobilized polynucleotide is immobilized by attachment to a solid support, and, through hybridization to the mediator polynucleotide, functions to immobilize the entire immobilized polynucleotide/target polynucleotide/probe polynucleotide "sandwich".

Abstract: A method for the preparation of a 3' end functionalized polynucleotide is disclosed. An amine-functionalized solid phase support is treated sequentially with an anhydride, then with an .omega.-hydroxylamine. A polynucleotide is chemically synthesized on the treated support and is subsequently cleaved therefrom by hydrolysis of the amide bonds. A polynucleotide having a 3' free primary amine is recovered for use in hybridization assays and other uses.

Abstract: DNA sequences encoding for novel human lymphoblastoid-leukocyte hybrid interferons. Methods of making nucleotide sequences comprising ribosome binding sites, promoter regions, and segments of human lymphoblastoid and leukocyte interferons are described. DNA sequences for hybrid interferons are cloned and expressed in plasmid vectors. Transformed microorganisms and novel lymphoblastoid-leukocyte polypeptide interferons having biological or immunological activity are disclosed.

Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences in excess of about 200 base pairs in length, which sequences may comprise entire structural genes. Novel sequences are prepared from two or more DNA subunits provided with terminal regions comprising restriction endonuclease cleavage sites facilitating insertion of subunits into a selected vector for purposes of amplification during the course of the total assembly process. The total, finally-assembled assembled sequences include at least one, and preferably two or more, unique restriction endonuclease cleavage site(s) at intermediate positions along the sequence, allowing for easy excision and replacement of subunits and the correspondingly facile preparation of multiple structural analogs of polypeptides coded for by the sequences.

Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences of up to about 200 base pairs, which sequences may comprise entire structural genes. Illustratively disclosed is the preparation and expression of manufactured genes, including fusion genes, capable of directing synthesis of human .beta.-endorphin and of proteins which differ from human .beta.-endorphin in terms of the identity or relative position of one or more amino acids. Manufactured genes preferably include codons selected from among alternative codons specifying the same amino acid on the basis of preferential expression in a projected host microorganism (e.g., E. coli) to be transformed.

Abstract: Illustrated is the preparation and expression of manufactured genes capable of directing synthesis of human immune and leukocyte interferons and of other biologically active proteinaceous products, which products differ from naturally-occurring forms in terms of the identity and/or relative position of one or more amino acids, and in terms of one or more biological and pharmacological properties but which substantially retain other such properties.