Isolation of Novel Transcribed Sequences from FRAXA Site via Exon Trapping
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摘要:
外显子捕获是近年发展起来的一种自基因组DNA分离表达序列的有效方法.我们采用以pSPL3为剪接载体的外显子捕获系统，从覆盖FRAXA位点的酵母人工染色体YAC209G4中分离到2个新外显子序列A91和D12.A91在人胎肝和骨骼肌文库中丰度较高，在肝、肾和骨髓文库中也有PCR扩增.而D12在所分析的8个cDNA文库中均未见PCR扩增产物.从骨骼肌文库中克隆到一段包含A91的315 bp的cDNA(AM4470).多种组织RNA印迹显示，AM4470与骨骼肌和心脏mRNA杂交，转录本长度均为2.8 kb.
Abstract:
Exon Trapping is a powerful method developed recently for isolating transcribed sequences from genomic DNA. Two novel exons, A91 and D12, were isolated from YAC209G4 spanning FRAXA site using the exon trapping system with pSPL3 as splicing vector. A91 was amplified strongly in fetal liver and skeletal muscle cDNA library and mildly in liver, kidney and bone marrow library, respectively. While D12 was failed to be detected from the 8 libraries. A fragment of 315 bp containing A91 has been cloned (AM4470) from the skeletal muscle cDNA library. A single transcript of 2.8 kb was detected from mRNA samples of human heart and skeletal muscle on multiple tissues Northern blot using AM4470 probe.
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