Su H, Lau Y F
Department of Medicine, VA Medical Center, University of California, San Francisco 94121.
Am J Hum Genet. 1993 Jan;52(1):24-38.
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.
通过运用一种涉及黏粒介导的基因转移、cDNA文库构建及分子特征分析技术的简单策略，我们确定了人类Y染色体上性别决定区Y（SRY）基因（即假定的睾丸决定因子（TDF）基因）的转录单元、结构组织及启动子序列。通过这种方法，分离出了携带人类SRY序列的重组黏粒，并将其转染至合适的组织培养细胞中。从转染细胞的聚腺苷酸加尾RNA构建的cDNA文库中分离出了重组cDNA克隆。对各个cDNA与基因组黏粒进行的比较研究提供了有关SRY基因及其mRNA组织的信息。结果表明，人类SRY基因是一个无内含子基因，产生1.1 kb的转录本，并且在其上游序列约310 bp处的转染细胞中具有启动子活性。
