Identification of a Ferritin Light Chain Pseudogene Near the Glycerol Kinase Locus in Xp21 by cDNA Amplification for Identification of Genomic Expressed Sequences

Weiwen Guo , Volker Adams , Jestina Mason , Edward R.B. McCabe
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引用次数: 1

Abstract

We used cDNA amplification for identification of genomic expressed sequences (CAIGES) to identify genes in the glycerol kinase region of the human X chromosome. During these investigations we identified the sequence for a ferritin light chain (FTL) pseudogene in this portion of Xp21. A human liver cDNA library was amplified by vector primers, labeled, and hybridized to Southern blots ofEcoRIdigested human genomic DNA from cosmids isolated from yeast artificial chromosomes in the glycerol kinase region of Xp21. A 3.1-kb restriction fragment hybridized with the cDNA library, was subcloned and sequenced, and a 440-bp intronless sequence was found with strong similarity to the FTL coding sequence. Therefore, the FTL pseudogene that had been mapped previously to Xp22.3–21.2 was localized specifically to the glycerol kinase region. The CAIGES method permits rapid screening of genomic material and will identify genomic sequences with similarities to genes expressed in the cDNA library used to probe the cloned genomic DNA, including pseudogenes.

利用cDNA扩增技术鉴定Xp21中甘油激酶位点附近铁蛋白轻链假基因的基因组表达序列
我们使用cDNA扩增技术鉴定基因组表达序列(CAIGES)来鉴定人类X染色体甘油激酶区域的基因。在这些研究中,我们在Xp21的这一部分中确定了铁蛋白轻链(FTL)假基因的序列。用载体引物扩增人肝脏cDNA文库,标记并杂交到ecoridigase人类基因组DNA的Southern blots上,这些DNA是从酵母人工染色体中分离出来的,位于Xp21的甘油激酶区域。将其与cDNA文库杂交得到的3.1 kb限制性内切片段进行亚克隆和测序,发现一个440-bp的无内含子序列与FTL编码序列具有较强的相似性。因此,之前定位于Xp22.3-21.2的FTL假基因特异性定位于甘油激酶区域。CAIGES方法允许快速筛选基因组物质,并将识别与cDNA文库中表达的基因相似的基因组序列,用于探测克隆的基因组DNA,包括假基因。
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