NDLI: Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins.

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Characterization of an essential Saccharomyces cerevisiae gene related to RNA processing: cloning of RNA1 and generation of a new allele with a novel phenotype.

Characterization of pp60src phosphorylation in vitro in Rous sarcoma virus-transformed cell membranes.

Membrane mutants of animal cells: rapid identification of those with a primary defect in glycosylation.

Cloning, sequencing, and functional analysis of oriL, a herpes simplex virus type 1 origin of DNA synthesis.

Nonrandom integration of human U4 RNA pseudogenes.

Ubiquitin is a heat shock protein in chicken embryo fibroblasts.

Stimulation of expression of a herpes simplex virus DNA-binding protein by two viral functions.

Site-specific DNA binding of nuclear factor I: analyses of cellular binding sites.

Suppression of tropomyosin synthesis, a common biochemical feature of oncogenesis by structurally diverse retroviral oncogenes.

Specific regulation of transcription of the discoidin gene family in Dictyostelium discoideum.

TU elements: a heterogeneous family of modularly structured eucaryotic transposons.

Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

Monoclonal proliferation of Friend murine leukemia virus-transformed myeloblastic cells occurs early in the leukemogenic process.

Purification and characterization of human H-ras proteins expressed in Escherichia coli.

Characterization, expression, and evolution of the mouse embryonic zeta-globin gene.

Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins.

Differential ability of a T-antigen transport-defective mutant of simian virus 40 to transform primary and established rodent cells.

Surface T-antigen expression in simian virus 40-transformed mouse cells: correlation with cell growth rate.

Low level of cellular protein phosphorylation by nontransforming overproduced p60c-src.

Preferential expression of the c-fps protein in chicken macrophages and granulocytic cells.

Overexpressed pp60c-src can induce focus formation without complete transformation of NIH 3T3 cells.

Nuclei of adenovirus 2-infected cells contain an RNA species that corresponds to an intron excised intact from mRNA precursors.

Processing of the precursor to a chloroplast ribosomal protein made in the cytosol occurs in two steps, one of which depends on a protein made in the chloroplast.

Regulation of protein synthesis factor EF-1 alpha in Mucor racemosus.

Alterations in chromatin structure associated with glucocorticoid-induced expression of endogenous mouse mammary tumor virus genes.

Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris.

Human cellular src gene: nucleotide sequence and derived amino acid sequence of the region coding for the carboxy-terminal two-thirds of pp60c-src.

A 12-O-tetradecanoylphorbol-13-acetate (TPA)-nonproliferative variant of 3T3 cells is resistant to TPA-enhanced gene amplification.

Bacteriophage lambda vector for transducing a cDNA clone library into mammalian cells.

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Novel chicken actin gene: third cytoplasmic isoform.

Ethidium bromide-induced loss of mitochondrial DNA from primary chicken embryo fibroblasts.

DNA primase-DNA polymerase alpha from simian cells: sequence specificity of initiation sites on simian virus 40 DNA.

Biochemical and immunological characterization of mutant L-M cells with altered levels of dibutyryl cyclic AMP-inducible alkaline phosphatase.

Gene transfer method for transient gene expression, stable transformation, and cotransformation of suspension cell cultures.

The simian virus 40 sequences between 0.169 and 0.423 map units are not essential to immortalize early-passage rat embryo cells.

Functional Analysis of the Transcription Control Region Located Within the Avian Retroviral Long Terminal Repeat

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Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins.

Content Provider	PubMed Central
Author	Alwine, J. C.
Abstract	The effects of trans-acting factors and transfected DNA stability on promoter activity were examined with chloramphenicol acetyl transferase (CAT) transient expression analysis. With cotransfection into CV-1P and HeLa cells, simian virus 40 T antigen, adenovirus E1a, and herpes-virus IE proteins were compared for their ability to trans-activate a variety of eucaryotic promoters constructed into CAT plasmids. T antigen and the IE protein were promiscuous activators of all the promoters tested [the simian virus 40 late promoter, the adenovirus E3 promoter, the alpha 2(I) collagen promoter, and the promoter of the Rous sarcoma virus long terminal repeat]. Conversely the E1a protein was specific, activating only the adenovirus E3 promoter and suppressing the basal activity of the other promoters. This specificity of activation by E1a contrasted with the high activity generated by all of the promoter-CAT plasmids when transfected into 293 cells, which endogenously produce E1a protein. Examination of transfected 293 cells determined that they stabilized much greater amounts of plasmid DNA than any other cells tested (CV-1P, COS, NIH-3T3, KB). Thus the high activity of nonadenovirus promoter-CAT plasmids in 293 cells results from the cumulative effect of basal promoter activity from a very large number of gene copies, not from E1a activation. This conclusion was supported by similar transfection analysis of KB cell lines which endogenously produce E1a protein. These cells stabilize plasmid DNA at a level comparable to that of CV-1P cells and, in agreement with the CV-1P cotransfection results, did not activate a nonadenovirus promoter-CAT plasmid. These results indicate that the stability of plasmid DNA must be considered when transient gene expression is being compared between cell lines. The use of relative plasmid copy numbers for the standardization of transient expression results is discussed.
Starting Page	1034
File Format	PDF
ISSN	10985549
e-ISSN	10985549
Journal	Molecular and Cellular Biology
Issue Number	5
Volume Number	5
Language	English
Publisher Date	1985-05-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Molecular Biology

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