Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1999-04-13
2000-08-01
Schwartzman, Robert A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 691, 4353201, 435325, 435455, 536 231, 536 241, C12Q 168, C12P 2100, C12N 1563, C12N 500, C07H 2104
Patent
active
060965053
ABSTRACT:
A noncloning method for expressing a gene of interest in a mammalian host cell is disclosed. The invention utilizes three basic individual elements: (1) a promoter element; (2) at least one gene of interest; and (3) a selectable marker cassette which includes in 5' to 3' order, an internal ribosome entry site ("IRES"), at least one gene coding for a selectable marker, and a transcription termination sequence. The three individual elements are cotransfected into a mammalian host cell where they become operably linked such that expression of the selectable marker gene(s) necessarily requires coexpression of the gene of interest.
REFERENCES:
patent: 4710463 (1987-12-01), Murray
Penolazzi et al. Direct transfection of polymerase chain reactoin generated DNA fragments into mammalian cells employing ethidium bromide inidcator and ultrafiltration. Anal. Biochem. vol. 248:190-193, Feb. 1997.
Campbell, et al. Homologous recombinatino involving small single-stranded oligonucleotides in human cells. The New Biologist. vol. 1(2):223-237, Nov. 1989.
Malpiece et al. Coloniy hybridization method for screening si situ of eukaryotic amplified genes. Experientia. Vo. 40:483-485, Mar. 1984.
Song et al. Effect or double-strand breaks on homologous recombination in mammalian cells and extracts. Mol. and Cellular Biol. vol. 5(12):3331-3336, Dec. 1985.
He et al. Construction of adenoviral and retroviral vectors coexpressing the genes encoding the hepatitis B surface antigen and B7-1 protein. Gene vol. 175:121-125, Nov. 1996.
Blazquez et al., "Mutations in the aphA-2 Gene of Transposon Tn5 Mapping Within the Regions Highly Conserved in Arminoglyoside-Phosphotransferases Strongly Reduce Aminoglycoside Resistance," Molecular Microbiology 5(6):1511-1518 (1991).
Escher and Schaffner, "Improved "Activator Trap" Method for the Isolation of Transcriptional Activation Domains From Random DNA Fragments," Biotechniques 1:848-854 (1996).
Gurtu et al., "IRES Bicistronic Expression Vectors for Efficient Creation of Stable Mammalian Cell Lines," Biochemical and Biophysical Research Communications 229:295-298 (1996).
Herlitschka et al., "Overexpression of Human Prothrombin in Permanent Cell Lines Using a Dominant Selection/Amplification Fusion Marker," Protein Expression and Purification 8:358-364 (1996).
Kaufman et al., "Improved Vectors for Stable Expression of Foreign Genes in Mammalian Cells by Use of the Untranslated Leader Sequence From EMC Virus," Nucleic Acids Research 19(16)4485-4490 (1991).
Kobayashi et al., "Improved Dicistronic mRNA Expression Vectors for Efficient Selection of Transfectants highly Expressing Foreign Genes," Bio Techniques 21:398-402 (1996).
Mosser et al., "Use of a Dicistronic Expression Cassette Encoding the Green Fluorescent Protein for the Screening and Selection of Cells Expressing Inducible Gene Products," Bio Techniques 22:150-161 (1997).
Penolazzi et al., "Direct Transfection of Polymerase Chain Reaction-Generated DNA Fragments into Mammalian Cells Employing Ethidium Bromide Indicator and Ultrafiltration," Analytical Biochemistry 248:190-193 (1997).
Rees et al., "Bicistronic Vector for the Creation of Stable Mamalian Cell Lines that Predisposes All Antibiotic-Resistant Cells to Express Recombinant Protein," Bio Techniques 20:102-110 (1996).
Dina Dino
Selby Mark
Thudium Kent B.
Blackburn Robert P.
Chiron Corporation
Harbin Alisa A.
Robins Roberta L.
Sandals William
LandOfFree
Noncloning technique for expressing a gene of interest does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Noncloning technique for expressing a gene of interest, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Noncloning technique for expressing a gene of interest will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-662350