Bio-technology General Corporation

Yissum Research Development Company of the Hebrew University of Jerusalem Ltd.

Other

Amos B Oppenheim

Marian Gorecki

Avigdor Levanon

Haim Aviv

Tikva Vogel

Jacob R Hartman

Menachem Zeevi

Elisha P Zeelon

Hilla Locker-Giladi

Rachel Oren

Hilla Locker-Galadi

Hilla Giladi

Idit Azar

Daniel Goldenberg

Ariella Oppenheim

Ziv Sandalon

Giladi Locker

Simi Kobi

Pinhas E Zeelon

Tikva Vogil

Simi Koby

The invention relates to constructs capable of infecting mammalian cells comprising at least one semi-purified or pure SV40 capsid protein and a constituent selected from the group consisting of an exogenous DNA, a vector comprising an exogenous DNA, an exogenous RNA, a vector comprising an exogenous RNA, an exogenous protein or peptide product, and antisense RNA, ribozyme RNA or any RNA or DNA which inhibits or prevents the expression of undesired protein(s) in the mammalian cell and optionally further comprising operatively linked regulatory elements sufficient for the expression and/or replication of the exogenous protein in a mammalian cell. The invention further relates to a method for the in vitro construction of SV40 virus or pseudovirus constructs according to the invention.

In vitro construction of SV40 viruses and pseudoviruses

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

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

The invention relates to an expression vector, the nucleic acid sequence of which comprises a promoter that is capable of controlling, when the vector is in a bacterial host and the temperature is lowered to below about 20.degree. C., the production of recombinant peptide or protein encoded by a gene contained within the vector. The invention also relates to host cells containing the vectors and to a method of producing a recombinant protein.

Vectors and transformed host cells for recombinant protein production at

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 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 for transcribing mRNA; and 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; a DNA sequence which contains an origin of replication capable of autonomous production in the host of at least 400 constitutive copies of the vector; and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host cell or the fragment cI.sup.434 on which is included the gene for the repressor protein and its associated promoter and operator. 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. Plasmids have been constructed from the vectors and used to produce bovine growth hormones.

Expression vectors containing lambdapl promoter, T.sub.1 T.sub.2 RRNA

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 bindig 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 fragment designated CI.sup.434 on which is included the gene for the repressor protein and its associated promoter and operator. Additionally, the vector may include a gene associated with a selectable or identifiable phenotypic trait which is manifested when the vector 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. Plasmids have been constructed from the vectors and used to produce bovine growth hormones.

Growing community of inventors

Jerusalem, Israel

Amos B Oppenheim