Abstract
We have developed an assay for specific genes in DNA based on transient expression. DNA prepared from patients with acute myeloblastic or acute lymphoblastic leukemia or from the continuous leukemic cell line HL60 was transferred to LTA cells; 48-72 hr later, these recipients expressed hemopoietic differentiation markers as detected by monoclonal antibodies against My-1 (granulocyte specific) and OK-T3 (T-cell specific) using immunofluorescence. The efficiency of transfer was dose and time dependent. We found that genes not expressed in the original cells were expressed after transfer by using this assay. Restriction enzyme analysis showed that My-1 was not expressed with DNA that had been incubated before transfer with either HindIII or Sal I but was present after digestion with either EcoRI or BamHI; after digestion of DNA with the same enzymes, OK-T3 expression was observed only in the HindIII-treated DNA. These studies indicate that DNA was transferred; this approach may provide an efficient method for detecting the activity of specific genes in the absence of selection.
MeSH Terms
Animals
Antibodies, Monoclonal
Cell Differentiation
Cells, Cultured
Cycloheximide/pharmacology
DNA Restriction Enzymes
DNA, Neoplasm/genetics
Dactinomycin/pharmacology
Fluorescent Antibody Technique
Kinetics
Leukemia, Lymphoid/genetics
Leukemia, Myeloid, Acute/genetics
Mice
Protein Biosynthesis/drug effects
Transcription, Genetic/drug effects
Transfection
Chemicals
Antibodies, Monoclonal
DNA, Neoplasm
Dactinomycin
Cycloheximide
DNA Restriction Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Chang L J
Gamble C L
Izaguirre C A
Minden M D
Mak T W
McCulloch E A
References (20)
20 references, click to expand
-
A new technique for the assay of infectivity of human adenovirus 5 DNA.
Virology. 1973 Apr;52(2):456-67
PMID: 4705382
-
Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis.
Biochemistry. 1973 Jul 31;12(16):3055-63
PMID: 4354250
-
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
J Mol Biol. 1975 Nov 5;98(3):503-17
PMID: 1195397
-
Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture.
Nature. 1977 Nov 24;270(5635):347-9
PMID: 271272
-
DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells.
Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373-6
PMID: 286319
-
Separation of blast cell and T-lymphocyte progenitors in the blood of patients with acute myeloblastic leukemia.
Blood. 1979 Jul;54(1):186-95
PMID: 312670
-
Characterisation of deletions which affect the expression of fetal globin genes in man.
Nature. 1979 Jun 14;279(5714):598-603
PMID: 450109
-
Transformation of mammalian cells with genes from procaryotes and eucaryotes.
Cell. 1979 Apr;16(4):777-85
PMID: 222468
-
The structure of the human beta-globin gene in beta-thalassaemia.
Nucleic Acids Res. 1979 Jun 25;6(8):2749-60
PMID: 461203
-
Monoclonal antibodies defining distinctive human T cell surface antigens.
Science. 1979 Oct 19;206(4416):347-9
PMID: 314668
-
DNA sequence variants in the G gamma-, A gamma-, delta- and beta-globin genes of man.
Cell. 1979 Sep;18(1):1-10
PMID: 509514
-
Passage of phenotypes of chemically transformed cells via transfection of DNA and chromatin.
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5714-8
PMID: 230490
-
Transforming activity of DNA of chemically transformed and normal cells.
Nature. 1980 Apr 3;284(5755):418-21
PMID: 7360280
-
Altering genotype and phenotype by DNA-mediated gene transfer.
Science. 1980 Sep 19;209(4463):1414-22
PMID: 7414320
-
Expression of a bacterial gene in mammalian cells.
Science. 1980 Sep 19;209(4463):1422-7
PMID: 6251549
-
High efficiency transformation by direct microinjection of DNA into cultured mammalian cells.
Cell. 1980 Nov;22(2 Pt 2):479-88
PMID: 6256082
-
Unique transforming gene in carcinogen-transformed mouse cells.
Nature. 1981 Feb 12;289(5798):607-9
PMID: 6258087
-
My-1, new myeloid-specific antigen identified by a mouse monoclonal antibody.
Blood. 1981 May;57(5):842-5
PMID: 6163493
-
Transforming activity of human tumor DNAs.
Proc Natl Acad Sci U S A. 1981 Feb;78(2):1181-4
PMID: 6940134
-
Efficient DNA transfection and rapid assay for thymidine kinase activity and viral antigenic determinants.
Somatic Cell Genet. 1981 Mar;7(2):161-70
PMID: 6170119