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

Journal Article · Mon Jul 01 04:00:00 EDT 1996 · Genomics

DOI:https://doi.org/10.1006/geno.1996.0344· OSTI ID:466040

Tanaka, Toshihiro ^[1]; Ogiwara, Atsushi; Uchiyama, Ikuo ^[2]

---

1. Univ. of Tokyo (Japan)
2. Univ. of Tokyo (Japan); and others

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.

• OSTI ID: 466040
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 1 Vol. 35; ISSN 0888-7543; ISSN GNMCEP
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
99 GENERAL AND MISCELLANEOUS
ANIMAL TISSUES
AUTOMATION
DATA BASE MANAGEMENT
DNA
DNA SEQUENCING
DNA-CLONING
EFFICIENCY
F CODES
GENE REGULATION
GENES
HEART
MITOCHONDRIA
TISSUE DISTRIBUTION