| Bicistronic vector for the creation of stable mammalian cell lines that predisposes all antibiotic-resistant cells to express recombinant protein. | |
| | |
MedLine Citation:
|
PMID: 8770413 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
We have developed an improved vector for the stable expression of recombinant protein in mammalian cells. In this vector, designated pCIN, both the recombinant cDNA and the neomycin phosphotransferase selection marker are transcribed from a single promoter element. To facilitate translation of the second open reading frame, the encephalomyocarditis virus internal ribosome entry site has been inserted into the expression cassette immediately before the start codon of this sequence. We report the use of this vector to generate stable cell lines expressing the human 5-HT1Da serotonin receptor and show that following transfection and clonal selection, all ten cell lines characterized express similar and high levels of receptor (1.5-11.9 pmol receptor/mg protein). Use of pCIN should permit the rapid and efficient production of stable mammalian cell lines for the characterization of recombinant protein, as this vector appears to predispose all transfected cells to express such protein. |
| | |
Authors:
|
S Rees; J Coote; J Stables; S Goodson; S Harris; M G Lee |
Related Documents
:
|
14769853 - Developmental separation of v(d)j recombinase expression and initiation of igh recombin... 15977183 - Cre-mediated site-specific recombination in zebrafish embryos. 16287483 - Identification of asymmetrically localized transcripts along the animal-vegetal axis of... |
Publication Detail:
|
Type: Technical Report |
Journal Detail:
|
Title: BioTechniques Volume: 20 ISSN: 0736-6205 ISO Abbreviation: BioTechniques Publication Date: 1996 Jan |
Date Detail:
|
Created Date: 1996-10-25 Completed Date: 1996-10-25 Revised Date: 2006-11-15 |
Medline Journal Info:
|
Nlm Unique ID: 8306785 Medline TA: Biotechniques Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 102-4, 106, 108-10 Citation Subset: IM |
Affiliation:
|
Receptor Systems Unit, Glaxo Wellcome Research and Development, Stevenage, Herts, UK. esr1353@ggr.co.uk@relay |
| Data Bank Information | |
Bank Name/Acc. No.:
|
GENBANK/U89672; U89673 |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Base Sequence Cell Line DNA Primers / genetics DNA, Complementary / genetics Drug Resistance / genetics* Gene Expression Genes / genetics* Genetic Vectors* Humans Kanamycin Kinase Molecular Sequence Data Phosphotransferases (Alcohol Group Acceptor) / genetics Plasmids / genetics Receptors, Serotonin / genetics Recombinant Proteins / genetics* Transfection |
| Chemical | |
Reg. No./Substance:
|
0/DNA Primers; 0/DNA, Complementary; 0/Receptors, Serotonin; 0/Recombinant Proteins; EC 2.7.1.-/Phosphotransferases (Alcohol Group Acceptor); EC 2.7.1.95/Kanamycin Kinase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Harvest protocol to reduce variability of soluble enzyme yield from cultured cells.
Next Document: Enhanced cytochemical detection of viral proteins and RNAs using double-sided labeling and light mic...