Nucleic Acids Research, 1983, Vol. 11, No. 20 7119-7136
© 1983
MOLECULAR BIOLOGY |
Plasmids for the cloning and expresion of full-length double-stranded cDNAs under control of the SV40 early or late gene promoter
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de 1'INSERM, Faculté de Médecine 11 Rue Humann, 67085 Strasbourg Cedex, France
Received August 30, 1983. Accepted September 27, 1983.
Okayama and Berg (1) have recently described a technique for the high efficiency cloning of full-length dscDNAs. We have constructed eukaryotic expression vectors compatible both with this technique (and with classical techniques) for dscDNA cloning. The vectors are such that recombinants obtained contain dscDNAs in the correct orientation downstream from a block of sequence comprising either the SV4O early or late gene promoter linked to a pair of splice sites from a rabbit ß-globin gene. A sequence encoding an SV40 polyadenylation site follows the dscDNA. We have used our vectors to make a library from chicken oviduct polyA(+) RNA using the Okayarma and Berg technique. Ovalbumin recombinants occur in the library at the expected frequency and a high proportion contain full length copies of the ovalbumin mRNA. However, a similar result was not obtained for conalbumin recombinants. When recombinants are introduced into eukaryotic cells by either calcium phosphate coprecipitation or protoplast fusion, expression of chicken ovalbumin or conalbumin may be detected by indirect immunofluorescence. Under optimal conditions (use of SV40 late promoter and cos 7 cells) ovalbumin protein could be detected when the ovalbumin recombinant was present in only 2 % of the protoplasts used for fusion. This suggests that colony banks obtained using our vectors could be screened in batches of 50 by protoplast fusion followed by a search for expression of a given protein using indirect immunofluorescence.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M.-B. Poirier, L. Laflamme, and M.-F. Langlois Identification and characterization of RanBPM, a novel coactivator of thyroid hormone receptors. J. Mol. Endocrinol., April 1, 2006; 36(2): 313 - 325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-B. Poirier, G. Hamann, M.-E. Domingue, M. Roy, T. Bardati, and M.-F. Langlois General Receptor for Phosphoinositides 1, a Novel Repressor of Thyroid Hormone Receptor Action that Prevents Deoxyribonucleic Acid Binding Mol. Endocrinol., August 1, 2005; 19(8): 1991 - 2005. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Oberste-Berghaus, K. Zanger, K. Hashimoto, R. N. Cohen, A. N. Hollenberg, and F. E. Wondisford Thyroid Hormone-independent Interaction between the Thyroid Hormone Receptor beta 2 Amino Terminus and Coactivators J. Biol. Chem., January 21, 2000; 275(3): 1787 - 1792. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. A. Rudolph-Owen, P. Cannon, and L. M. Matrisian Overexpression of the Matrix Metalloproteinase Matrilysin Results in Premature Mammary Gland Differentiation and Male Infertility Mol. Biol. Cell, February 1, 1998; 9(2): 421 - 435. [Abstract] [Full Text]
|
|
|
|
 |
|
 An Altered Intron Inhibits Synthesis of the Acetylcholine Receptor {alpha}-Subunit in the Paralyzed Zebrafish Mutant nic1 Genetics, January 1, 1998; 148(1): 361 - 372.
|
|
|
|
|
|
M.-F. Langlois, K. Zanger, T. Monden, J. D. Safer, A. N. Hollenberg, and F. E. Wondisford A Unique Role of the beta -2 Thyroid Hormone Receptor Isoform in Negative Regulation by Thyroid Hormone. MAPPING OF A NOVEL AMINO-TERMINAL DOMAIN IMPORTANT FOR LIGAND-INDEPENDENT ACTIVATION J. Biol. Chem., October 3, 1997; 272(40): 24927 - 24933. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. N. Hollenberg, T. Monden, and F. E. Wondisford Ligand-independent and -dependent Functions of Thyroid Hormone Receptor Isoforms Depend upon Their Distinct Amino Termini J. Biol. Chem., June 16, 1995; 270(24): 14274 - 14280. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C Glineur, M Zenke, H Beug, and J Ghysdael Phosphorylation of the v-erbA protein is required for its function as an oncogene. Genes & Dev., October 1, 1990; 4(10): 1663 - 1676. [Abstract] [PDF]
|
|
|
|
|
|
C Rochette-Egly, C Fromental, and P Chambon General repression of enhanson activity by the adenovirus-2 E1A proteins. Genes & Dev., January 1, 1990; 4(1): 137 - 150. [Abstract] [PDF]
|
|
|
|
 |
|
 B. Harris, J. Robishaw, S. Mumby, and A. Gilman Molecular cloning of complementary DNA for the alpha subunit of the G protein that stimulates adenylate cyclase Science, September 20, 1985; 229(4719): 1274 - 1277. [Abstract] [PDF]
|
|