This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Okayama, H Articles by Berg, P Search for Related Content PubMed PubMed Citation Articles by Okayama, H Articles by Berg, P

A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells.

ABSTRACT

This paper describes a plasmid vector for cloning cDNAs in Escherichia coli; the same vector also promotes expression of the cDNA segment in mammalian cells. Simian virus 40 (SV40)-derived DNA segments are arrayed in the pcD vector to permit transcription, splicing, and polyadenylation of the cloned cDNA segment. A DNA fragment containing both the SV40 early region promoter and two introns normally used to splice the virus 16S and 19S late mRNAs is placed upstream of the cDNA cloning site to ensure transcription and splicing of the cDNA transcripts. An SV40 late region polyadenylation sequence occurs downstream of the cDNA cloning site, so that the cDNA transcript acquires a polyadenylated 3' end. By using pcD-alpha-globin cDNA as a model, we confirmed that the alpha-globin transcript produced in transfected cells is initiated correctly, spliced at either of the two introns, and polyadenylated either at the site coded in the cDNA segment or at the distal SV40 polyadenylation signal. A cDNA clone library constructed with mRNA from SV40-transformed human fibroblasts and this vector (about 1.4 X 10(6) clones) yielded full-length cDNA clones that express hypoxanthine-guanine phosphoribosyltransferase (Jolly et al., Proc. Natl. Acad. Sci. U.S.A., in press).

This article has been cited by other articles:

• Huang, H. W., Bi, W., Jenkins, G. N., Alcorn, J. L. (2008). Glucocorticoid Regulation of Human Pulmonary Surfactant Protein-B mRNA Stability Involves the 3'-Untranslated Region. Am. J. Respir. Cell Mol. Bio. 38: 473-482 [Abstract] [Full Text]  
• Huang, X.-P., Ellis, J. (2007). Mutational Disruption of a Conserved Disulfide Bond in Muscarinic Acetylcholine Receptors Attenuates Positive Homotropic Cooperativity between Multiple Allosteric Sites and Has Subtype-Dependent Effects on the Affinities of Muscarinic Allosteric Ligands. Mol. Pharmacol. 71: 759-768 [Abstract] [Full Text]  
• Munakata, Y., Saito-Ito, T., Kumura-Ishii, K., Huang, J., Kodera, T., Ishii, T., Hirabayashi, Y., Koyanagi, Y., Sasaki, T. (2005). Ku80 autoantigen as a cellular coreceptor for human parvovirus B19 infection. Blood 106: 3449-3456 [Abstract] [Full Text]  
• Ostedgaard, L. S., Rokhlina, T., Karp, P. H., Lashmit, P., Afione, S., Schmidt, M., Zabner, J., Stinski, M. F., Chiorini, J. A., Welsh, M. J. (2005). A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia. Proc. Natl. Acad. Sci. USA 102: 2952-2957 [Abstract] [Full Text]  
• Gage, R. M., Matveeva, E. A., Whiteheart, S. W., von Zastrow, M. (2005). Type I PDZ Ligands Are Sufficient to Promote Rapid Recycling of G Protein-coupled Receptors Independent of Binding to N-Ethylmaleimide-sensitive Factor. J. Biol. Chem. 280: 3305-3313 [Abstract] [Full Text]  
• Cao, T. T., Brelot, A., von Zastrow, M. (2005). The Composition of the {beta}-2 Adrenergic Receptor Oligomer Affects Its Membrane Trafficking after Ligand-Induced Endocytosis. Mol. Pharmacol. 67: 288-297 [Abstract] [Full Text]  
• Tanowitz, M., von Zastrow, M. (2002). Ubiquitination-independent Trafficking of G Protein-coupled Receptors to Lysosomes. J. Biol. Chem. 277: 50219-50222 [Abstract] [Full Text]  
• Bannister, R. A., Melliti, K., Adams, B. A. (2002). Reconstituted Slow Muscarinic Inhibition of Neuronal (CaV1.2c) L-Type Ca2+ Channels. Biophys. J 83: 3256-3267 [Abstract] [Full Text]  
• Amendt, C., Mann, A., Schirmacher, P., Blessing, M. (2002). Resistance of keratinocytes to TGF{beta}-mediated growth restriction and apoptosis induction accelerates re-epithelialization in skin wounds. J. Cell Sci. 115: 2189-2198 [Abstract] [Full Text]  
• Ohara, O., Temple, G. (2001). Directional cDNA library construction assisted by the in vitro recombination reaction. Nucleic Acids Res 29: e22-e22 [Abstract] [Full Text]  
• Dennehy, K. M., Ferris, W. F., Veenstra, H., Zuckerman, L. A., Killeen, N., Beyers, A. D. (2001). Determination of the tyrosine phosphorylation sites in the T cell transmembrane glycoprotein CD5. Int Immunol 13: 149-156 [Abstract] [Full Text]  
• Gunning, P., Ferguson, V, Brennan, K., Hardeman, E. (2001). Alpha-skeletal actin induces a subset of muscle genes independently of muscle differentiation and withdrawal from the cell cycle. J. Cell Sci. 114: 513-524 [Abstract]  
• Carstens, C., Newman, D. K., Bohlen, H., Konig, A., Koch, N. (2000). Invariant chains with the class II binding site replaced by a sequence from influenza virus matrix protein constrain low-affinity sequences to MHC II presentation. Int Immunol 12: 1561-1568 [Abstract] [Full Text]  
• NEBERT, D. W., DALTON, T. P., STUART, G. W., CARVAN, M. J. III (2000). "Gene-Swap Knock-in" Cassette in Mice to Study Allelic Differences in Human Genes. Ann. N. Y. Acad. Sci. 919: 148-170 [Abstract] [Full Text]  
• Tsao, P. I., von Zastrow, M. (2000). Type-specific Sorting of G Protein-coupled Receptors after Endocytosis. J. Biol. Chem. 275: 11130-11140 [Abstract] [Full Text]  
• Shishido-Hara, Y., Hara, Y., Larson, T., Yasui, K., Nagashima, K., Stoner, G. L. (2000). Analysis of Capsid Formation of Human Polyomavirus JC (Tokyo-1 Strain) by a Eukaryotic Expression System: Splicing of Late RNAs, Translation and Nuclear Transport of Major Capsid Protein VP1, and Capsid Assembly. J. Virol. 74: 1840-1853 [Abstract] [Full Text]  
• Vickery, R. G., von Zastrow, M. (1999). Distinct Dynamin-dependent and -independent Mechanisms Target Structurally Homologous Dopamine Receptors to Different Endocytic Membranes. J. Cell Biol. 144: 31-43 [Abstract] [Full Text]  
• Ito, A., Morii, E., Maeyama, K., Jippo, T., Kim, D.-K., Lee, Y.-M., Ogihara, H., Hashimoto, K., Kitamura, Y., Nojima, H. (1998). Systematic Method to Obtain Novel Genes That Are Regulated by mi Transcription Factor: Impaired Expression of Granzyme B and Tryptophan Hydroxylase in mi/mi Cultured Mast Cells. Blood 91: 3210-3221 [Abstract] [Full Text]  
• Payen, E., Verkerk, T., Michalovich, D., Dreyer, S. D., Winterpacht, A., Lee, B., De Zeeuw, C. I., Grosveld, F., Galjart, N. (1998). The Centromeric/Nucleolar Chromatin Protein ZFP-37 May Function to Specify Neuronal Nuclear Domains. J. Biol. Chem. 273: 9099-9109 [Abstract] [Full Text]  
• Siebenkotten, I. M., Carstens, C., Koch, N. (1998). Identification of a Sequence That Mediates Promiscuous Binding of Invariant Chain to MHC Class II Allotypes. J. Immunol. 160: 3355-3362 [Abstract] [Full Text]  
• Kimura, M., Kaneko, I., Komiyama, M., Takatsuki, A., Koshino, H., Yoneyama, K., Yamaguchi, I. (1998). Trichothecene 3-O-Acetyltransferase Protects Both the Producing Organism and Transformed Yeast from Related Mycotoxins. CLONING AND CHARACTERIZATION OF Tri101. J. Biol. Chem. 273: 1654-1661 [Abstract] [Full Text]  
• Liu, Y., Bohn, S. A., Sherley, J. L. (1998). Inosine-5'-Monophosphate Dehydrogenase Is a Rate-determining Factor for p53-dependent Growth Regulation. Mol. Biol. Cell 9: 15-28 [Abstract] [Full Text]  
• Jiang, Q., Guo, D., Lee, B. X., Van Rhee, A. M., Kim, Y.-C., Nicholas, R. A., Schachter, J. B., Harden, T. K., Jacobson, K. A. (1997). A Mutational Analysis of Residues Essential for Ligand Recognition at the Human P2Y1 Receptor. Mol. Pharmacol. 52: 499-507 [Abstract] [Full Text]  
• Shofuda, K.-i., Yasumitsu, H., Nishihashi, A., Miki, K., Miyazaki, K. (1997). Expression of Three Membrane-type Matrix Metalloproteinases (MT-MMPs) in Rat Vascular Smooth Muscle Cells and Characterization of MT3-MMPs with and without Transmembrane Domain. J. Biol. Chem. 272: 9749-9754 [Abstract] [Full Text]  
• Hayami, K., Fukuta, D., Nishikawa, Y., Yamashita, Y., Inui, M., Ohyama, Y., Hikida, M., Ohmori, H., Takai, T. (1997). Molecular Cloning of a Novel Murine Cell-surface Glycoprotein Homologous to Killer Cell Inhibitory Receptors. J. Biol. Chem. 272: 7320-7327 [Abstract] [Full Text]  
• Koishi, R., Kawashima, I., Yoshimura, C., Sugawara, M., Serizawa, N. (1997). Cloning and Characterization of a Novel Oxidoreductase KDRF from a Human Bone Marrow-derived Stromal Cell Line KM-102. J. Biol. Chem. 272: 2570-2577 [Abstract] [Full Text]  
• Zhao, J., Paganini, L., Mucke, L., Gordon, M., Refolo, L., Carman, M., Sinha, S., Oltersdorf, T., Lieberburg, I., McConlogue, L. (1996). beta -Secretase Processing of the beta -Amyloid Precursor Protein in Transgenic Mice Is Efficient in Neurons but Inefficient in Astrocytes. J. Biol. Chem. 271: 31407-31411 [Abstract] [Full Text]  
• Mikulits, W., Hengstschläger, M., Sauer, T., Wintersberger, E., Müllner, E. W. (1996). Overexpression of Thymidine Kinase mRNA Eliminates Cell Cycle Regulation of Thymidine Kinase Enzyme Activity. J. Biol. Chem. 271: 853-860 [Abstract] [Full Text]  
• Saga, Y, Hata, N, Kobayashi, S, Magnuson, T, Seldin, M., Taketo, M. (1996). MesP1: a novel basic helix-loop-helix protein expressed in the nascent mesodermal cells during mouse gastrulation. Development 122: 2769-2778 [Abstract]  
• Kim, J., Wess, Jür., Rhee, A. M. v., Schöneberg, T., Jacobson, K. A. (1995). Site-directed Mutagenesis Identifies Residues Involved in Ligand Recognition in the Human A[IMAGE] Adenosine Receptor. J. Biol. Chem. 270: 13987-13997 [Abstract] [Full Text]  
• Lorenz, M, Jung, S, Radbruch, A (1995). Switch transcripts in immunoglobulin class switching. Science 267: 1825-1828 [Abstract]  
• Flynn, D. C., Koay, T. C., Humphries, C. G., Guappone, A. C. (1995). AFAP-120. J. Biol. Chem. 270: 3894-3899 [Abstract] [Full Text]  
• Burstein, E. S., Spalding, T. A., Hill-Eubanks, D., Brann, M. R. (1995). Structure-Function of Muscarinic Receptor Coupling to G Proteins. J. Biol. Chem. 270: 3141-3146 [Abstract] [Full Text]  
• Miyata, T, Takeda, J, Iida, Y, Yamada, N, Inoue, N, Takahashi, M, Maeda, K, Kitani, T, Kinoshita, T (1993). The cloning of PIG-A, a component in the early step of GPI-anchor biosynthesis. Science 259: 1318-1320 [Abstract]  
• Sandri-Goldin, R M, Mendoza, G E (1992). A herpesvirus regulatory protein appears to act post-transcriptionally by affecting mRNA processing.. Genes Dev. 6: 848-863 [Abstract]  
• Chan, A., Rubin, J., Bottaro, D., Hirschfield, D., Chedid, M, Aaronson, S. (1991). Identification of a competitive HGF antagonist encoded by an alternative transcript. Science 254: 1382-1385 [Abstract]  
• Miki, T, Fleming, T., Bottaro, D., Rubin, J., Ron, D, Aaronson, S. (1991). Expression cDNA cloning of the KGF receptor by creation of a transforming autocrine loop. Science 251: 72-75 [Abstract]  
• Richardson, H E, Stueland, C S, Thomas, J, Russell, P, Reed, S I (1990). Human cDNAs encoding homologs of the small p34Cdc28/Cdc2-associated protein of Saccharomyces cerevisiae and Schizosaccharomyces pombe.. Genes Dev. 4: 1332-1344 [Abstract]  
• Sisodia, S., Koo, E., Beyreuther, K, Unterbeck, A, Price, D. (1990). Evidence that beta-amyloid protein in Alzheimer's disease is not derived by normal processing. Science 248: 492-495 [Abstract]  
• Choi, K., Kim, H., Lee, S., Moon, K., Sim, S., Kim, J., Chung, H., Rhee, S. (1990). Molecular cloning and expression of a complementary DNA for inositol 1,4,5-trisphosphate 3-kinase. Science 248: 64-66 [Abstract]  
• Speck, N A, Renjifo, B, Golemis, E, Fredrickson, T N, Hartley, J W, Hopkins, N (1990). Mutation of the core or adjacent LVb elements of the Moloney murine leukemia virus enhancer alters disease specificity.. Genes Dev. 4: 233-242 [Abstract]  
• Rommens, J., Iannuzzi, M., Kerem, B, Drumm, M., Melmer, G, Dean, M, Rozmahel, R, Cole, J., Kennedy, D, Hidaka, N, al., e. (1989). Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245: 1059-1065 [Abstract]  
• Kasid, U, Pfeifer, A, Brennan, T, Beckett, M, Weichselbaum, R., Dritschilo, A, Mark, G. (1989). Effect of antisense c-raf-1 on tumorigenicity and radiation sensitivity of a human squamous carcinoma. Science 243: 1354-1356 [Abstract]  
• Jaskulski, D, deRiel, J., Mercer, W., Calabretta, B, Baserga, R (1988). Inhibition of cellular proliferation by antisense oligodeoxynucleotides to PCNA cyclin. Science 240: 1544-1546 [Abstract]  
• Goyert, S., Ferrero, E, Rettig, W., Yenamandra, A., Obata, F, Le Beau, M. (1988). The CD14 monocyte differentiation antigen maps to a region encoding growth factors and receptors. Science 239: 497-500 [Abstract]  
• Kasid, U, Pfeifer, A, Weichselbaum, R., Dritschilo, A, Mark, G. (1987). The raf oncogene is associated with a radiation-resistant human laryngeal cancer. Science 237: 1039-1041 [Abstract]  
• Ohtsubo, M, Kai, R, Furuno, N, Sekiguchi, T, Sekiguchi, M, Hayashida, H, Kuma, K, Miyata, T, Fukushige, S, Murotsu, T (1987). Isolation and characterization of the active cDNA of the human cell cycle gene (RCC1) involved in the regulation of onset of chromosome condensation.. Genes Dev. 1: 585-593 [Abstract]  
• Bonner, T., Buckley, N., Young, A., Brann, M. (1987). Identification of a family of muscarinic acetylcholine receptor genes. Science 237: 527-532 [Abstract]  
• Wong, P M, Chung, S W, Nienhuis, A W (1987). Retroviral transfer and expression of the interleukin-3 gene in hemopoietic cells.. Genes Dev. 1: 358-365 [Abstract]  
• Ono, Y, Kikkawa, U, Ogita, K, Fujii, T, Kurokawa, T, Asaoka, Y, Sekiguchi, K, Ase, K, Igarashi, K, Nishizuka, Y (1987). Expression and properties of two types of protein kinase C: alternative splicing from a single gene. Science 236: 1116-1120 [Abstract]  
• Zuber, M., Simpson, E., Waterman, M. (1986). Expression of bovine 17 alpha-hydroxylase cytochrome P-450 cDNA in nonsteroidogenic (COS 1) cells. Science 234: 1258-1261 [Abstract]  
• Yamamoto, T, Bishop, R., Brown, M., Goldstein, J., Russell, D. (1986). Deletion in cysteine-rich region of LDL receptor impedes transport to cell surface in WHHL rabbit. Science 232: 1230-1237 [Abstract]  
• Arya, S., Guo, C, Josephs, S., Wong-Staal, F (1985). Trans-activator gene of human T-lymphotropic virus type III (HTLV-III). Science 229: 69-73 [Abstract]  
• Neumann, J., Schach, N., Koch, N. (2001). Glycosylation Signals That Separate the Trimerization from the MHC Class II-binding Domain Control Intracellular Degradation of Invariant Chain. J. Biol. Chem. 276: 13469-13475 [Abstract] [Full Text]  
• Guittet, O., Hakansson, P., Voevodskaya, N., Fridd, S., Graslund, A., Arakawa, H., Nakamura, Y., Thelander, L. (2001). Mammalian p53R2 Protein Forms an Active Ribonucleotide Reductase in Vitro with the R1 Protein, Which Is Expressed Both in Resting Cells in Response to DNA Damage and in Proliferating Cells. J. Biol. Chem. 276: 40647-40651 [Abstract] [Full Text]  
• Gage, R. M., Kim, K.-A., Cao, T. T., von Zastrow, M. (2001). A Transplantable Sorting Signal That Is Sufficient to Mediate Rapid Recycling of G Protein-coupled Receptors. J. Biol. Chem. 276: 44712-44720 [Abstract] [Full Text]