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5187263 Expression of biologically active PDGE analogs in eucaryotic cells
PATENT ABSTRACTS lysis against pure water removes salts. Repeated lyophilization removes excess water and cow centrates single protamines and protamine-like proteins. The mixture may then be reconstituted with 5~ weight/volume heterologous or homoIogous DNA, in order to shield from charge toxicity. Crude mixtures may be produced by precipitating the supcruate of uitracentrifugation in pure water, followed by ultracentrifugation to sediment in solids. Lyophilization then removes any water from the damp solids. The crude solids are suitable for oral use, especially if utilized in gelatin capsules. Sterile filtration to injection quality aqueous form. Following isolation of the protamiue-DNA complex, encapsulation of the prepared solid or aqueous protamine-DNA complexes in a specific carrier substance may be accomplished, depending upon the target tissue for the protamine. Several encapsulation carders are known from prior art literature, such as, for example, liposomes and nanoparticles. The protamiue-DNA complexes of the present invention are useful in inhibiting tumor growth, among other uses.
5187262 CDNA ENCODING A POLYPEPTIDE INCLUDING HEVEIN SEQUENCE
A
Natasha V Raikhei, Willem F Broekaert, NamHal Chua, Anil Kush assigned to Board of Trustees operating Michigan State University A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed olignnucleotides corresponding to two regions of hevein as primers and a Hevea braeiliensis latex cDNA library as a template. HEVI is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence~f 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79~0 homologous to the carboxyl-terminal region of woundinducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the
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protein of the present invention and maltose binding protein produced by the E. coli.
5187263 EXPRESSION OF BIOLOGICALLY ACTIVE PDGE ANALOGS IN EUCARYOTIC CELLS Mark J Murray, James D Kelly assigned to ZymoGenetics Inc Methods for expressing a variety of biologically active PDGF analogs in eucaryotic cells are disclosed. The methods generally comprise introducing into a eucaryotic host cell a DNA construct capable of directing the expression and secretion of biologically active PDGF analogs in eucaryotic cells. The DNA construct contains a transcriptional promoter followed downstream by a suitable DNA sequence. The DNA sequence may encode a protein substantially homologous to the A-chain or the B-chain of PDGF, or a portion thereof, or an A-B heterodimer. In addition, a portion of the DNA sequence may encode at least a portion of the Achain, while another portion encodes at least a portion of the B-chain of PDGF. Eucaryotic cells transformed with these DNA constructs are also disclosed. Methods of promoting the growth of mammalian cells, comprising incubating the cells with a biologically active PDGF analog expressed by a eucaryotic host cell transformed with such a DNA construct, are also disclosed.
51~2~ PLANT PROTEINS, PROMOTERS, CODING SEQUENCES AND USE Luea Comai, Ann J Koning assigned to Calgene Inc Novel nucleic acid sequences, constructs employing such sequences, transgenic plant cells and plants are provided employing sequences associated with tomato heat shock protein 80.5 ( hspg0 ). Nucleic acid sequences obtainable from the use of hspg0 as a probe in other plant species are also disclosed. In particular, the 5' noncoding region of tomato hsp80 or other plant promoters obtainable therefrom, especially to provide differentially specific initiation of transcription of a desired nucleic acid sequence of interest. Also provided are putative sequences believed to represent Scaffold Attachment Regions (SAR) of the plant hsp80 gene.
5187262 CDNA ENCODING A POLYPEPTIDE INCLUDING HEVEIN SEQUENCE
A
Natasha V Raikhei, Willem F Broekaert, NamHal Chua, Anil Kush assigned to Board of Trustees operating Michigan State University A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed olignnucleotides corresponding to two regions of hevein as primers and a Hevea braeiliensis latex cDNA library as a template. HEVI is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence~f 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79~0 homologous to the carboxyl-terminal region of woundinducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the
107
protein of the present invention and maltose binding protein produced by the E. coli.
5187263 EXPRESSION OF BIOLOGICALLY ACTIVE PDGE ANALOGS IN EUCARYOTIC CELLS Mark J Murray, James D Kelly assigned to ZymoGenetics Inc Methods for expressing a variety of biologically active PDGF analogs in eucaryotic cells are disclosed. The methods generally comprise introducing into a eucaryotic host cell a DNA construct capable of directing the expression and secretion of biologically active PDGF analogs in eucaryotic cells. The DNA construct contains a transcriptional promoter followed downstream by a suitable DNA sequence. The DNA sequence may encode a protein substantially homologous to the A-chain or the B-chain of PDGF, or a portion thereof, or an A-B heterodimer. In addition, a portion of the DNA sequence may encode at least a portion of the Achain, while another portion encodes at least a portion of the B-chain of PDGF. Eucaryotic cells transformed with these DNA constructs are also disclosed. Methods of promoting the growth of mammalian cells, comprising incubating the cells with a biologically active PDGF analog expressed by a eucaryotic host cell transformed with such a DNA construct, are also disclosed.
51~2~ PLANT PROTEINS, PROMOTERS, CODING SEQUENCES AND USE Luea Comai, Ann J Koning assigned to Calgene Inc Novel nucleic acid sequences, constructs employing such sequences, transgenic plant cells and plants are provided employing sequences associated with tomato heat shock protein 80.5 ( hspg0 ). Nucleic acid sequences obtainable from the use of hspg0 as a probe in other plant species are also disclosed. In particular, the 5' noncoding region of tomato hsp80 or other plant promoters obtainable therefrom, especially to provide differentially specific initiation of transcription of a desired nucleic acid sequence of interest. Also provided are putative sequences believed to represent Scaffold Attachment Regions (SAR) of the plant hsp80 gene.