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Cloning and Sequencing a cDNA which is Related to D13S25 and might be Linked to the Gene Locus for Wilson's Disease
以分子生物學方法於人類染色體威爾遜病病因區內選殖出一段靠近D13S25之基因的互補DNA並訂定其DNA次序
摘要
本研究的目的在試圖以染色體的長臂上q14帶上的一個2.2 Kb的DNA片段H2-42為探針篩選人的染色體DNA庫（genomic library）以分離出一個靠近威爾遜（Wilson's disease）致病相關帶的基因。篩選結果得到一個強陽性的溶菌斑，此溶菌斑的λ-dash含有一段約13.6Kb長的染色體DNA。可用Hind III限制酶切割成5.3Kb，2.7Kb，2.2Kb，1.5Kb，1.2Kb及0.7Kb片段。其中2.2Kb， 1.5Kb及1.2Kb 片段被用來進一步篩選胎盤及Y79互補基因庫（cDNA library）。其中1.5Kb DNA 片段自胎盤互補基因庫中找出兩株陽性的溶菌斑。此1.6Kb互補DNA的次序經訂定後發現並未含有長段可轉讀性的DNA或poly A的片段。
並列摘要
A 2.2 Kb genomic fragment H2-42 (DI3S25) was used as a probe to screen λ-dash human genomic library with an attempt to isolate a gene which is located in the associated range of the gene locus for Wilson's disease. Single positive plague was isolated. This clone contained a 13.6 Kb genomic insert which could be cut by Hind III into 5.3 Kb, 2.7 Kb , 2.2Kb , 1.5 Kb , 1.2 Kb and 0.7 Kb fragments. Those fragments with a size of 2.2 Kb, 1.5 Kb and 1.2 Kb were used to screen human placenta and Y-79 cDNA libraries. Two independent clones were identified from placenta cDNA library by 1.5 Kb fragment. Both clones contained the same 1.6 Kb cDNA insert with an internal EcoR I cutting site. This cDNA was sequenced and it contained neither open reading frame nor poly A segment.
延伸閱讀
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