IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES BY RESTRICTION ENDONUCLEASE-BASED GENE-EXPRESSION FINGERPRINTING

被引：56

作者：

IVANOVA, NB

BELYAVSKY, AV

机构：

[1] RUSSIAN ACAD SCI,VA ENGELHARDT MOLEC BIOL INST,MOSCOW 117984,RUSSIA

[2] NEW YORK BLOOD CTR,NEW YORK,NY 10021

来源：

NUCLEIC ACIDS RESEARCH | 1995年 / 23卷 / 15期

关键词：

D O I：

10.1093/nar/23.15.2954

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：2954 / 2958

页数：5

共 17 条

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