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PMID: 6275384 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Detection of genes coding for human differentiation markers by their transient expression after DNA transfer.

Chang LJ, Gamble CL, Izaguirre CA, Minden MD, Mak TW, McCulloch EA

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
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20 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1982-01-00
Pages
146-50
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC345679
Subset
IM
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