Bio-technology General Corporation

Yeda Research and Develpment & Co. Ltd

Ferring International Center Sa.

Marian Gorecki

Amos B Oppenheim

Avigdor Levanon

Haim Aviv

Tikva Vogel

Jacob R Hartman

Menachem Zeevi

Elisha P Zeelon

Rachel Oren

Hilla Locker-Galadi

Alexander Rich

Meir Wilchek

Hilla Locker-Giladi

Amnon Gonenne

Simona Mendelovitz

Simona Mendelovitch

Netta Fulga

Clemenceau T Acquaye

Pinhas E Zeelon

Arie Rosner

Eliyahu Keshet

Tikva Vogil

The present invention relates to strains of,adapted to produce bovine growth hormone, being the result of extraction of RNA from bovine pituitaries, transcription of DNA on such RNA templates, splicing of DNA into plasmids, insertion of spliced plasmid into microorganisms, and further subjecting the microorganisms to selection and isolating the colonies producing the desired fused or modified growth hormone and to modified bovine growth hormone obtained by the cultivation of such modified,strains and recovery of the desired product.

Production of bovine growth hormone by microorganisms

An improved vector upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of a desired gene inserted into the vector and production of polypeptide encoded by the gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the desired gene capable of binding to ribosomes within the host cell; an ATG initiation codon or a DNA sequence which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a restriction enzyme site for inserting the desired gene into the vector in phase with the ATG initiation codon; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the vector is present in the host cell.

Expression vectors for enhanced production of polypeptides, plasmids

Improved methods for production of recombinant human superoxide dismutase and analogs thereof have been developed. Such superoxide dismutase or analogs may be used to catalyze the reduction of superoxide radicals thereby preventing or treating injury resulting from the presence of such superoxide radicals.

Therapeutic SOD compositions and uses thereof

An improved and efficient process for the production of recombinant human insulin by folding of a proinsulin hybrid polypeptide is provided.

Refolding of proinsulins without addition of reducing agents

The subject invention provides a method of producing enzymatically active CPB which comprises treating a recombinant cell containing DNA encoding ProCPB, so that the DNA directs expression of the ProCPB, recovering from the cell the ProCPB so expressed, treating the recovered ProCPB under conditions permitting folding of the ProCPB, subjecting the folded ProCPB to enzymatic cleavage to produce enzymatically active CPB and purifying the enzymatically active CPB.

Production of enzymatically active recombinant carboxypeptidase B

An improved vector upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzyme site for inserting the gene in phase with the ATG codon; a T.sub.1 T.sub.2 rRNA transcription termination sequence; an origin of replication and a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host. The distance between the 3' end of the P.sub.L O.sub.L promoter and operator sequence and the 5' end of the N utilization site is less than about 80 base pairs and the distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Plasmids have been constructed from the vectors and used to produce bovine, chicken and porcine growth hormones, human apolipoprotein E and human superoxide dismutase.

Expression vectors containing .lambda.P.sub.L promoter and T.sub.1

A plasmid for the production of superoxide dismutase or an analog thereof which upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of DNA encoding superoxide dismutase and production of superoxide dismutase. The plasmid is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the gene encoding superoxide dismutase capable of binding to ribosomes within the host cell; an ATG initiation codon; a restriction enzyme site for inserting the gene encoding superoxide dismutase into the plasmid in phase with the ATG initiation codon; a gene encoding superoxide dismutase; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the plasmid is present in the host cell.

Bacterial expression of superoxide dismutase

Plasmids for production of human growth hormone or polypeptide analog

Expression vectors containing lambdapl promoter and T.sub.1 T.sub.2 rRNA

A novel analog of human superoxide dismutase which comprises the non-acetylated, non-glycosylated form of natural human superoxide dismutase having the ser-met amino acid sequence attached to the N-terminus and a mixture comprising this analog and non-acetylated, non-glycosylated superoxide dismutase having an amino acid sequence identical to that of natural human superoxide dismutase are provided. Methods of catalyzing a chemical reaction using the novel analog and mixture are provided.

Sod polypeptide analogs

An improved plasmid for the production of superoxide dismutase on an analog thereof which upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a gene encoding superoxide dismutase or the analog. The plasmid is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon; a second restriction enzyme site; a gene encoding superoxide dismutase or the analog; an origin of replication and a gene associated with a selectable or identifiable phenotypic trait manifested when the plasmid is present in the host. The distance between the 3' end of P.sub.L O.sub.L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Host vector systems have been constructed from these plasmids and used to produce human superoxide dismutase and analogs thereof. Such SOD or analogs may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and reduce spinal cord injury.

Therapeutic compositions comprising a mixture of human CuZn superoxide

A plasmid for the production of porcine growth hormone or an analog thereof which upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of DNA encoding porcine growth hormone and production of porcine growth hormone or analogs thereof. The plasmid is a double-stranded DNA molecule which includes in 5' to 3' the following: a DNA sequence which contains the promoter and operator PLOL from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the gene encoding porcine growth hormone capable of binding to ribosomes within the host cell; an ATG initiation codon; a restriction enzyme site for inserting the gene encoding porcine growth hormone into the plasmid in phase with the ATG initiation codon; a gene encoding porcine growth hormone; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the plasmid is present in the host cell.

Bacterial expression of porcine growth hormone

Plasmids for production of met-asp-gln bovine growth hormone, hosts

An improved plasmid for the production of superoxide dismutase which upon introduction into a host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a gene encoding superoxide dismutase. The plasmid which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon; a second restriction enzyme site; a gene encoding superoxide dismutase; an origin of replication and a gene associated with a selectable or identifiable phenotypic trait manifested when the plasmid is present in the host. The distance between the 3' end of P.sub.L O.sub.L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Host vector systems have been constructed from these plasmids and used to produce human superoxide dismutase and analogs thereof. Such SOD or analogs may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and reduce spinal cord injury.

Plasmids for expression of human superoxide dismutase (SOD) analogs

An improved plasmid for the production of superoxide dismutase which upon introduction into a host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a gene encoding superoxide dismutase. The plasmid includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon; a second restriction enzyme site; a gene encoding superoxide dismutase; an origin of replication and a gene associated with a selectable or identifiable phenotypic trait manifested when the plasmid is present in the host. The distance between the 3' end of P.sub.L O.sub.L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Host vector systems have been constructed from these plasmids and used to produce human superoxide dismutase and analogs thereof. Such SOD or analogs may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and reduce spinal cord injury.

Growing community of inventors

Rehovot, Israel

Marian Gorecki