skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Construction of a normalized directionally cloned cDNA library from adult heart and analysis of 3040 clones by partial sequencing

Abstract

Large-scale sequencing of clones from cDNA libraries derived from specific tissues is a rapid and efficient way of discovering novel genes expressed in those tissues. However, because the heart is continually contracting and relaxing, it strongly expresses muscle contractile genes and/or mitochondrial genes, a bias that reduces the efficiency of this method. To improve the efficiency of identifying novel genes expressed in the heart, we constructed a normalized directionally cloned cDNA library from adult heart and partially sequenced 3040 clones. Comparisons of these sequence data with known, 23.4% were identical or almost identical to human expressed sequence tags (ESTs), 14.2% bore no significant homology to any sequences in the database, and 1.2% represented repetitive sequences. The remaining 4.1% showed some homology with known gene, and Northern blot analysis of several clones in this category revealed that most of them were expressed mainly in the heart and skeletal muscle. After redundancy was excluded, the 3040 clones accounted for 1395 distinctive ESTs, 446 of which exhibited no match to any known sequence. Our results suggest that our normalized library is less redundant than standard libraries and is a useful resource for cataloging genes expressed in the heart. 19 refs., 1 fig., 2more » tabs.« less

Authors:
 [1]; ;  [2]
  1. Univ. of Tokyo (Japan)
  2. Univ. of Tokyo (Japan); and others
Publication Date:
OSTI Identifier:
466040
Resource Type:
Journal Article
Journal Name:
Genomics
Additional Journal Information:
Journal Volume: 35; Journal Issue: 1; Other Information: PBD: 1 Jul 1996
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; 99 MATHEMATICS, COMPUTERS, INFORMATION SCIENCE, MANAGEMENT, LAW, MISCELLANEOUS; DNA SEQUENCING; EFFICIENCY; DATA BASE MANAGEMENT; F CODES; AUTOMATION; DNA; DNA-CLONING; GENE REGULATION; GENES; TISSUE DISTRIBUTION; HEART; ANIMAL TISSUES; MITOCHONDRIA

Citation Formats

Tanaka, Toshihiro, Univ. of Tokyo School of Medicine, Hongo, Ogiwara, Atsushi, and Uchiyama, Ikuo. Construction of a normalized directionally cloned cDNA library from adult heart and analysis of 3040 clones by partial sequencing. United States: N. p., 1996. Web. doi:10.1006/geno.1996.0344.
Tanaka, Toshihiro, Univ. of Tokyo School of Medicine, Hongo, Ogiwara, Atsushi, & Uchiyama, Ikuo. Construction of a normalized directionally cloned cDNA library from adult heart and analysis of 3040 clones by partial sequencing. United States. doi:10.1006/geno.1996.0344.
Tanaka, Toshihiro, Univ. of Tokyo School of Medicine, Hongo, Ogiwara, Atsushi, and Uchiyama, Ikuo. Mon . "Construction of a normalized directionally cloned cDNA library from adult heart and analysis of 3040 clones by partial sequencing". United States. doi:10.1006/geno.1996.0344.
@article{osti_466040,
title = {Construction of a normalized directionally cloned cDNA library from adult heart and analysis of 3040 clones by partial sequencing},
author = {Tanaka, Toshihiro and Univ. of Tokyo School of Medicine, Hongo and Ogiwara, Atsushi and Uchiyama, Ikuo},
abstractNote = {Large-scale sequencing of clones from cDNA libraries derived from specific tissues is a rapid and efficient way of discovering novel genes expressed in those tissues. However, because the heart is continually contracting and relaxing, it strongly expresses muscle contractile genes and/or mitochondrial genes, a bias that reduces the efficiency of this method. To improve the efficiency of identifying novel genes expressed in the heart, we constructed a normalized directionally cloned cDNA library from adult heart and partially sequenced 3040 clones. Comparisons of these sequence data with known, 23.4% were identical or almost identical to human expressed sequence tags (ESTs), 14.2% bore no significant homology to any sequences in the database, and 1.2% represented repetitive sequences. The remaining 4.1% showed some homology with known gene, and Northern blot analysis of several clones in this category revealed that most of them were expressed mainly in the heart and skeletal muscle. After redundancy was excluded, the 3040 clones accounted for 1395 distinctive ESTs, 446 of which exhibited no match to any known sequence. Our results suggest that our normalized library is less redundant than standard libraries and is a useful resource for cataloging genes expressed in the heart. 19 refs., 1 fig., 2 tabs.},
doi = {10.1006/geno.1996.0344},
journal = {Genomics},
number = 1,
volume = 35,
place = {United States},
year = {1996},
month = {7}
}