Rommens J M, Durocher F, McArthur J, Tonin P, LeBlanc J F, Allen T, Samson C, Ferri L, Narod S, Morgan K
Department of Genetics, Hospital for Sick Children, University of Toronto, Ontario, Canada.
Genomics. 1995 Aug 10;28(3):530-42. doi: 10.1006/geno.1995.1185.
A detailed transcription map of the 320-kb region containing the HSD17B locus on chromosome 17 was generated. Thirty unique cDNA fragments, retrieved following the hybridization of immobilized YACs to primary pools of cDNAs prepared from RNA of mammary gland, ovary, placenta, and the Caco-2 cell line, were aligned into 10 transcription units by physical mapping and hybridization to RNAs of a series of tissues. The cDNAs were then further characterized by sequencing and used to screen mammary gland cDNA libraries. Fragments corresponding to the broadly expressed gamma-tubulin and Ki antigen genes were identified. A full-length cDNA clone encoding a 117-amino-acid protein homologous to the rat ribosomal protein L34 was isolated. Portions of genes with restricted patterns of expression were also obtained, including the previously characterized HSD17B1. One new gene, for which a full-length cDNA was isolated, was found to have an interesting tissue-specific pattern of expression with abundant mRNA in both the colon and the testis and in the mammary carcinoma cell line BT-474. This contrasted with the barely detectable level observed in several tissues including normal mammary gland. Of the five additional transcription units identified, one showed no similarity, two showed identity to human expressed sequences, and two displayed similarity to genes of animal species by amino acid alignment. These latter cDNA clones include potential homologues of a rat nuclear tyrosine phosphatase and of a factor of Drosophila that is known to be involved in the negative regulation of transcription of segment identity genes.
构建了包含17号染色体上HSD17B基因座的320 kb区域的详细转录图谱。通过物理图谱分析以及与一系列组织的RNA杂交,将从乳腺、卵巢、胎盘和Caco-2细胞系的RNA制备的cDNA一级文库与固定化YAC杂交后获得的30个独特cDNA片段排列成10个转录单位。然后通过测序对这些cDNA进行进一步表征,并用于筛选乳腺cDNA文库。鉴定出了与广泛表达的γ-微管蛋白和Ki抗原基因相对应的片段。分离出一个编码与大鼠核糖体蛋白L34同源的117个氨基酸的蛋白质的全长cDNA克隆。还获得了表达模式受限的部分基因,包括先前已表征的HSD17B1。发现了一个新基因,其全长cDNA已被分离,该基因具有有趣的组织特异性表达模式,在结肠、睾丸和乳腺癌细胞系BT-474中均有丰富的mRNA表达。这与在包括正常乳腺在内的几种组织中观察到的几乎检测不到的水平形成对比。在鉴定出的另外五个转录单位中,一个没有相似性,两个与人表达序列相同,两个通过氨基酸比对显示与动物物种的基因相似。后两个cDNA克隆包括大鼠核酪氨酸磷酸酶和果蝇中已知参与节段身份基因转录负调控的一个因子的潜在同源物。