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© 1992 Oxford University Press

RESEARCH-ARTICLE

Molecular isolation and characterization of an expressed gene from the human Y chromosome

Julie S. Zhang, Teresa L. Yang-Feng1, Ulrich Muller2, T.K. Mohandas3, Pieter J. de Jong4 and Yan-Fai Chris Lau*

Division of Cell and Developmental Genetics, VA Medical Center, University of California San Francisco, 4150 Clement Street, San Francisco, CA 94121 1Department of Human Genetics, School of Medicine, Yale University New Haven, CT 06510 2Genetics Division, Children's Hospital Medical Center Boston, MA 02115 3Division of Medical Genetics, Harbor-UCLA Medical Center Torrance, CA 90509 4Human Genome Center, Lawrence Livermore National Laboratory Livermore, CA 94550, USA

* To whom correspondence should be addressed

Received July 23, 1992; Revised October 16, 1992; Accepted October 16, 1992

Using a positional cloning approach, we have isolated an expressed gene from a flow-sorted Y chromosome cosmid library. The isolation of this gene was based on the identification of the Y-231 cosmid that contains CpG rich sequences (HTF islands) in its human insert. The Y-231 cosmid was capable of detecting a 1.3 kb transcript In poly (A) RNA samples from human testis. Several cDNA clones were isolated from a human testis cDNA library constructed in {lambda}gtl0. In addition, DNA-mediated gene transfer and restriction enzyme mapping experiments demonstrated that two functional transcriptional units are present within the Y-231 cosmid. DNA sequencing analysis showed that the largest cDNA clone contains 1075 bp of unique sequence and a poly (A) track at the 3' end of the corresponding mRNA. An open reading frame of 762 bp that encodes a predicted protein of 253 amino acids with a calculated molecular weight of 28.9 kD was identified. The Y-231 structural gene encompasses approximately 2.7 kb of genomic sequence and contains six exons that are interrupted by five introns. The Y-231 gene shares very high (97%) identity at the DNA level to a previously described Y-spedfic gene, testis specific protein V-encoded (TSPY) gene, suggesting the possibility that these two genes are related, if not identical. However, the TSPY gene has been postulated to be intronless. Further PCR and RT-PCR analyses of these two genes and their transcripts have provided evidence supporting the hypothesis that they are the same gene and are members of a Y-speclfic repeated gene family containing intronic sequences. The Y-231 (TSPY) gene is conserved In the male genome and expressed in the testis of the chimpanzee, suggesting that it may play an important role in the physiology of this organ in man and the great ape.


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