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IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES BY RESTRICTION ENDONUCLEASE-BASED GENE-EXPRESSION FINGERPRINTING
Author(s): IVANOVA NB, BELYAVSKY AV
Source: NUCLEIC ACIDS RESEARCH    Volume: 23    Issue: 15    Pages: 2954-2958    Published: AUG 11 1995  
Times Cited: 55     References: 17     
Abstract: A novel method for identification of differentially expressed genes has been developed, It is based on the consecutive restriction digestions of 3' terminal cDNA fragments to produce a fingerprint of gene expression, cDNA molecules are synthesized using a biotinylated oligo(dT) primer, digested with a frequently cutting restriction endonuclease and the 3'-terminal restriction fragments are isolated using streptavidin microbeads, After amplification by PCR, cDNA fragments are immobilized again on streptavidin beads, radiolabeled and treated sequentially with a set of restriction endonucleases, The products of individual enzymatic reactions from two or more different RNA populations are resolved by polyacrylamide gel electrophoresis and compared to reveal differentially expressed genes. This strategy enabled us to identify and clone the fragments of five genes expressed differentially in murine thymus and spleen, One of the genes was found to encode terminal deoxynucleotidyl transferase; others are apparently previously unknown genes.
Document Type: Article
Language: English
Addresses:
1. RUSSIAN ACAD SCI, VA ENGELHARDT MOLEC BIOL INST, MOSCOW 117984, RUSSIA
2. NEW YORK BLOOD CTR, NEW YORK, NY 10021 USA
Publisher: OXFORD UNIV PRESS UNITED KINGDOM, WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP
Subject Category: Biochemistry & Molecular Biology
IDS Number: RR361
ISSN: 0305-1048
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