Isolation and characterization of 21 novel expressed DNA sequences from the distal region of human chromosome 4p

Journal Article · Fri Jul 15 04:00:00 EDT 1994 · Genomics

DOI:https://doi.org/10.1006/geno.1994.1388· OSTI ID:249987

Ishida, Yoshikazu; Hadano, Shinji; Nagayama, Tomiko ^[1]

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1. Tokai Univ. School of Medicine, Kanagawa (Japan); and others

The authors have established an approach to the isolation of expressed DNA sequences from a defined region of the human chromosome. The method relies on the direct screening of cDNA libraries using pooled single-copy microclones generated by a laser chromosome microdissection in conjunction with a single unique primer polymerase chain reaction (SUP-PCR) procedure. They applied this method to the distal region of human chromosome 4p (4p15-4pter), which contains the Huntington disease (HD) and the Wolf-Hirschhorn syndrome (WHS) loci. Twenty-one nonoverlapping and region-specific cDNA clones encoding novel genes were isolated in this manner. Ten of 21 clones were subregionally assigned to 4p16.1-4pter, and the remainder mapped to the region proximal to 4p16.1. Northern blot and reverse transcription followed by the PCR (RT-PCR) analysis revealed that 16 of these 21 clones detected transcripts in total RNA from human tissues. The method is applicable to other chromosomal regions and is a powerful approach to the isolation of region-specific cDNA clones. 44 refs., 3 figs., 3 tabs.

• OSTI ID: 249987
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 2 Vol. 22; ISSN 0888-7543; ISSN GNMCEP
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
DNA HYBRIDIZATION
GENETIC MAPPING
HUMAN CHROMOSOMES
OPTIMIZATION