Advertisement
No access
Report

Differential Display of Eukaryotic Messenger RNA by Means of the Polymerase Chain Reaction

Science
14 Aug 1992
Vol 257, Issue 5072
pp. 967-971

Abstract

Effective methods are needed to identify and isolate those genes that are differentially expressed in various cells or under altered conditions. This report describes a method to separate and clone individual messenger RNAs (mRNAs) by means of the polymerase chain reaction. The key element is to use a set of oligonucleotide primers, one being anchored to the polyadenylate tail of a subset of mRNAs, the other being short and arbitrary in sequence so that it anneals at different positions relative to the first primer. The mRNA subpopulations defined by these primer pairs were amplified after reverse transcription and resolved on a DNA sequencing gel. When multiple primer sets were used, reproducible patterns of amplified complementary DNA fragments were obtained that showed strong dependence on sequence specificity of either primer.

Get full access to this article

View all available purchase options and get full access to this article.

References

ADAMS, M.D., SEQUENCE IDENTIFICATION OF 2,375 HUMAN BRAIN GENES, NATURE 355: 632 (1992).
Alberts, B., Molecular Biology of the Cell (1989).
BAND, V, TUMOR PROGRESSION IN 4 MAMMARY EPITHELIAL-CELL LINES DERIVED FROM THE SAME PATIENT, CANCER RESEARCH 50: 7351 (1990).
Bothwell, A., Methods for Cloning and Analysis of Eukaryotic Genes (1990).
COPPOCK, D.L., CONTROL OF THYMIDINE KINASE MESSENGER-RNA DURING THE CELL-CYCLE, MOLECULAR AND CELLULAR BIOLOGY 7: 2925 (1987).
Croy, R. G., Proceedings of the National Academy of Sciences of the United States of America 80: 4699 (1983).
HOFBAUER, R, CELL-CYCLE REGULATED SYNTHESIS OF STABLE MOUSE THYMIDINE KINASE MESSENGER-RNA IS MEDIATED BY A SEQUENCE WITHIN THE CDNA, NUCLEIC ACIDS RESEARCH 15: 741 (1987).
JACKSON, R.J., DO THE POLY(A) TAIL AND 3' UNTRANSLATED REGION CONTROL MESSENGER-RNA TRANSLATION, CELL 62: 15 (1990).
KEYOMARSI, K, SYNCHRONIZATION OF TUMOR AND NORMAL-CELLS FROM G1 TO MULTIPLE CELL-CYCLES BY LOVASTATIN, CANCER RESEARCH 51: 3602 (1991).
KHAN, A.S., EFFICIENT DOUBLE-STRANDED SEQUENCING OF CDNA CLONES CONTAINING LONG POLY(A) TAILS USING ANCHORED POLY(DT) PRIMERS, NUCLEIC ACIDS RESEARCH 19: 1715 (1991).
LEE, S.W., POSITIVE SELECTION OF CANDIDATE TUMOR-SUPPRESSOR GENES BY SUBTRACTIVE HYBRIDIZATION, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 88: 2825 (1991).
LEWIN, B, DRIVING THE CELL-CYCLE - M-PHASE KINASE, ITS PARTNERS, AND SUBSTRATES, CELL 61: 743 (1990).
LIANG, P, unpublished data.
LIN, P.F., MOLECULAR-CLONING AND STRUCTURAL-ANALYSIS OF MURINE THYMIDINE KINASE GENOMIC AND CDNA SEQUENCES, MOLECULAR AND CELLULAR BIOLOGY 5: 3149 (1985).
MANIATIS, T, REGULATION OF INDUCIBLE AND TISSUE-SPECIFIC GENE-EXPRESSION, SCIENCE 236: 1237 (1987).
MASIAKOWSKI, P, CLONING OF CDNA SEQUENCES OF HORMONE-REGULATED GENES FROM THE MCF-7 HUMAN-BREAST CANCER CELL-LINE, NUCLEIC ACIDS RESEARCH 10: 7895 (1982).
OFARRELL, P.H., HIGH-RESOLUTION 2-DIMENSIONAL ELECTROPHORESIS OF PROTEINS, JOURNAL OF BIOLOGICAL CHEMISTRY 250: 4007 (1975).
PARDEE, A.B., G1 EVENTS AND REGULATION OF CELL-PROLIFERATION, SCIENCE 246: 603 (1989).
SU, J.Y., MOLECULAR-CLONING AND EXPRESSION OF THE HUMAN DEOXYTHYMIDYLATE KINASE GENE IN YEAST, NUCLEIC ACIDS RESEARCH 19: 823 (1991).
Varmus, H., Oncogenes and the Molecular Origins of Cancer: 3 (1989).
WILLIAMS, JGK, DNA POLYMORPHISMS AMPLIFIED BY ARBITRARY PRIMERS ARE USEFUL AS GENETIC-MARKERS, NUCLEIC ACIDS RESEARCH 18: 6531 (1990).

(0)eLetters

eLetters is a forum for ongoing peer review. eLetters are not edited, proofread, or indexed, but they are screened. eLetters should provide substantive and scholarly commentary on the article. Embedded figures cannot be submitted, and we discourage the use of figures within eLetters in general. If a figure is essential, please include a link to the figure within the text of the eLetter. Please read our Terms of Service before submitting an eLetter.

Log In to Submit a Response

No eLetters have been published for this article yet.

Information & Authors

Information

Published In

Science
Volume 257 | Issue 5072
14 August 1992

Submission history

Published in print: 14 August 1992

Permissions

Request permissions for this article.

Authors

Affiliations

Peng Liang
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, and Division of Cell Growth and Regulation, Dana-Farber Cancer Institute, Boston, MA 02115.
Arthur B. Pardee
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, and Division of Cell Growth and Regulation, Dana-Farber Cancer Institute, Boston, MA 02115.

Metrics & Citations

Metrics

Article Usage

Altmetrics

Citations

Cite as

Export citation

Select the format you want to export the citation of this publication.

Cited by

  1. Noxa, a BH3-Only Member of the Bcl-2 Family and Candidate Mediator of p53-Induced Apoptosis, Science, 288, 5468, (1053-1058), (2021)./doi/10.1126/science.288.5468.1053
    Abstract
  2. Single-Cell Gene Expression Profiling, Science, 297, 5582, (836-840), (2021)./doi/10.1126/science.1072241
    Abstract
  3. A Family of cAMP-Binding Proteins That Directly Activate Rap1, Science, 282, 5397, (2275-2279), (1998)./doi/10.1126/science.282.5397.2275
    Abstract
  4. Induction of Gene Expression in After Pilus-Mediated Adherence, Science, 273, 5279, (1234-1236), (1996)./doi/10.1126/science.273.5279.1234
    Abstract
  5. Alternatives to 35S as a Label for the Differential Display of Eukaryotic Messenger RNA, Science, 267, 5201, (1186-1187), (1995)./doi/10.1126/science.7855603
    Abstract
  6. Cloning of Immunoglobulin-Superfamily Members Associated with HLA-C and HLA-B Recognition by Human Natural Killer Cells, Science, 268, 5209, (405-408), (1995)./doi/10.1126/science.7716543
    Abstract
  7. Serial Analysis of Gene Expression, Science, 270, 5235, (484-487), (1995)./doi/10.1126/science.270.5235.484
    Abstract
  8. Response: Alternatives to 35S as a Label for the Differential Display of Eukaryotic Messenger RNA, Science, 267, 5201, (1186-1187), (1995)./doi/10.1126/science.267.5201.1186.b
    Abstract
Loading...

View Options

Check Access

Log in to view the full text

AAAS ID LOGIN

AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.

Log in via OpenAthens.
Log in via Shibboleth.

More options

Purchase digital access to this article

Download and print this article for your personal scholarly, research, and educational use.

Purchase this issue in print

Buy a single issue of Science for just $15 USD.

View options

PDF format

Download this article as a PDF file

Download PDF

Media

Figures

Multimedia

Tables

Share

Share

Share article link

Share on social media