Nucleic Acids Research, 1993, Vol. 21, No. 14 3269-3275
© 1993
METHODS |
Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization
Division of Cell Growth and Regulation, Dana-Farber Cancer Institute and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School Boston, MA 02115, USA
*To whom correspondence should be addressed
Received March 3, 1993. Revised June 9, 1993. Accepted June 9, 1993.
Differential display has been developed as a tool to detect and characterize altered gene expression in eukaryotic cells. The basic principle Is to systematically amplify messenger RNA5 and then distribute theIr 3' termini on a denaturing polyacrylamide gel. Here we provide methodological details and examine in depth the specificity, sensitivity and reproducibIlity of the method. We show that the number of anchored 01190- dT primers can be reduced from twelve to four that are degenerate at the penultimate base from the 3' end. We also demonstrate that using optimized conditions described here, multiple RNA samples from related cells can be displayed simultaneously. Therefore process-specific rather than cell-specific genes could be more accurately identified. These results enable further streamlining of the technique and make It readily applicable to a broad spectrum of biological systems.
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M. M. Montano and B. S. Katzenellenbogen The quinone reductase gene: A unique estrogen receptor-regulated gene that is activated by antiestrogens PNAS, March 18, 1997; 94(6): 2581 - 2586. [Abstract] [Full Text] [PDF]
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J. M. Babity, J. N. Armstrong, J.-C. L. Plumier, R. W. Currie, and H. A. Robertson A novel seizure-induced synaptotagmin gene identified by differential display PNAS, March 18, 1997; 94(6): 2638 - 2641. [Abstract] [Full Text] [PDF]
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H. Simon, R Kittappa, P. Khan, C Tsilfidis, R. Liversage, and S Oppenheimer A novel family of T-box genes in urodele amphibian limb development and regeneration: candidate genes involved in vertebrate forelimb/hindlimb patterning Development, January 4, 1997; 124(7): 1355 - 1366. [Abstract] [PDF]
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K. Kuno, N. Kanada, E. Nakashima, F. Fujiki, F. Ichimura, and K. Matsushima Molecular Cloning of a Gene Encoding a New Type of Metalloproteinase-disintegrin Family Protein with Thrombospondin Motifs as an Inflammation Associated Gene J. Biol. Chem., January 3, 1997; 272(1): 556 - 562. [Abstract] [Full Text] [PDF]
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M. Shen, H Wang, and P Leder A differential display strategy identifies Cryptic, a novel EGF-related gene expressed in the axial and lateral mesoderm during mouse gastrulation Development, January 1, 1997; 124(2): 429 - 442. [Abstract] [PDF]
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E. A. Ariazi and M. N. Gould Identifying Differential Gene Expression in Monoterpene-treated Mammary Carcinomas Using Subtractive Display J. Biol. Chem., November 15, 1996; 271(46): 29286 - 29294. [Abstract] [Full Text] [PDF]
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C.-M. Hsieh, M. Yoshizumi, W. O. Endege, C.-J. Kho, M. K. Jain, S. Kashiki, R. d. l. Santos, W.-S. Lee, M. A. Perrella, and M.-E. Lee APEG-1, a Novel Gene Preferentially Expressed in Aortic Smooth Muscle Cells, Is Down-regulated by Vascular Injury J. Biol. Chem., July 19, 1996; 271(29): 17354 - 17359. [Abstract] [Full Text] [PDF]
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E. Hu, P. Liang, and B. M. Spiegelman AdipoQ Is a Novel Adipose-specific Gene Dysregulated in Obesity J. Biol. Chem., May 3, 1996; 271(18): 10697 - 10703. [Abstract] [Full Text] [PDF]
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Y. Ueda and G. Chaudhuri Differential Expression of B1-containing Transcripts in Leishmania-exposed Macrophages J. Biol. Chem., June 16, 2000; 275(25): 19428 - 19432. [Abstract] [Full Text] [PDF]
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