Investigation of gyrate atrophy using a cDNA clone for human ornithine aminotransferase.
Investigation of gyrate atrophy using a cDNA clone for human ornithine aminotransferase.
复制标题
使用人鸟氨酸转氨酶 cDNA 克隆研究回旋萎缩。
DOI：
10.1089/dna.1.1986.5.493
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Gusella,JF
中科院分区：
文献类型：
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作者：
Ramesh,V;Shaffer,MM;Allaire,JM;Shih,VE;Gusella,JF
脉络膜和视网膜回旋萎缩是一种常染色体隐性遗传疾病，与鸟氨酸转氨酶 (OAT) 活性降低或缺失相关。为了在分子水平上解决 OAT 的缺陷，我们从人肝脏中克隆了编码 OAT 前体的 mRNA 的 cDNA。该克隆包含 1317 个核苷酸的完整编码区以及 44 个核苷酸的 5' 和 654 个核苷酸的 3' 非翻译序列。当用于探测基因组 DNA 时，OAT cDNA 没有在脑回萎缩患者中检测到任何基因缺失或重排的证据。该 cDNA 与肝脏、成纤维细胞和淋巴母细胞中的 2.15 kb RNA 杂交。在 7 名脑回萎缩患者的成纤维细胞和/或淋巴母细胞中，该 mRNA 的大小和大致数量没有改变，这些患者的 OAT 活性降低了 25 至 100 倍。我们的结果表明，这些个体的缺陷可能是由 mRNA 中的细微序列改变引起的，但这种改变并不影响其表观大小。
Gyrate atrophy of the choroid and retina is an autosomal recessive disease associated with reduced or absent ornithine aminotransferase (OAT) activity. To approach the defect in OAT at the molecular level, we have cloned a cDNA for the mRNA encoding the OAT precursor from human liver. The clone contains the complete coding region of 1317 nucleotides along with 44 nucleotides of 5′ and 654 nucleotides of 3′ untranslated sequences. When used to probe genomic DNA, the OAT cDNA did not detect any evidence of gene deletion or rearrangement in patients with gyrate atrophy. The cDNA hybridizes to a 2.15-kb RNA species in liver, fibroblasts, and lymphoblasts. The size and approximate amount of this mRNA is not altered in fibroblasts and/or lymphoblasts of seven gyrate atrophy patients who display a 25- to 100-fold reduction in OAT activity. Our results suggest the defect in these individuals may be caused by a subtle sequence alteration in the mRNA that does not affect its apparent size.