Identification of the transcriptional unit, structural organization, and promoter sequence of the human sex-determining region Y (SRY) gene, using a reverse genetic approach

• Published in: American journal of human genetics Vol. 52; no. 1; pp. 24 - 38
• Main Authors: HUA SU, LAU, Y.-F. C
• Format: Journal Article
• Language: English
• Published: Chicago, IL University of Chicago Press 1993
• Subjects: Amino Acid Sequence; Animals; Base Sequence; BASIC BIOLOGICAL SCIENCES; Biological and medical sciences; cDNA; Cell Line; CHROMOSOMES; Classical genetics, quantitative genetics, hybrids; CLONING; CONSTRUCTION; COSMIDS; DNA; DNA HYBRIDIZATION; DNA-Binding Proteins - genetics; DNA-CLONING; Female; Fundamental and applied biological sciences. Psychology; GENE REPRESSORS; GENES; Genetics of eukaryotes. Biological and molecular evolution; HETEROCHROMOSOMES; Human; HUMAN CHROMOSOMES; HUMAN Y CHROMOSOME; Humans; HYBRIDIZATION; Introns; LIBRARIES; Male; man; Mice; Molecular Sequence Data; Nuclear Proteins; NUCLEIC ACIDS; nucleotide sequence; ORGANIC COMPOUNDS; Promoter Regions, Genetic; promoters; RNA; SEX; sex determination; Sex Determination Analysis; Sex-Determining Region Y Protein; SRY gene; Transcription Factors; Transcription, Genetic; Y Chromosome; Y CHROMOSOME 550400 > Genetics; Human; Nucleotide sequence; Transcription; Sex determination; Cosmid; Gene organization; Y-Chromosome; Recombinant DNA; Technique; Molecular biology; Sexual differentiation
• ISSN: 0002-9297; 1537-6605

Using a simple strategy involving cosmid-mediated gene transfer, cDNA library construction, and molecular characterization techniques, we have determined the transcriptional unit, structural organization, and promoter sequence of the human sex-determining region Y (SRY) gene, the putative testis-determining factor (TDF) gene on the human Y chromosome. By this approach, a recombinant cosmid harboring the human SRY sequence was isolated and transfected to appropriate tissue-cultured cells. Recombinant cDNA clones were isolated from a cDNA library constructed from poly (A) + RNA of the transfected cells. Comparative studies between the respective cDNAs and the genomic cosmid have provided information regarding the organization of the SRY gene and its mRNAs. The results indicate that the human SRY gene is an intronless gene, produces transcripts of 1.1 kb, and possesses promoter activities in the transfected cells at approximately 310 bp of its upstream sequences.