论文部分内容阅读
目的 预测HumCyr6 1基因的变异剪接转录本 ,并通过分析其在不同组织中的表达谱进行验证 ,以便初步探讨HumCyr6 1基因的功能。 方法 应用计算机辅助的电子杂交步移策略 ,通过对比查询数据库返回的ESTs序列和先前克隆的HumCyr6 1cDNA序列 ,预测了HumCyr6 1基因的变异剪接转录本。将HumCyr6 1基因片段制备探针后与含有人体 8个组织mRNA的MTNI膜杂交 ,进行表达谱分析。结果 计算机预测HumCyr6 1基因存在 3个潜在的变异剪接位点 ,暗示该基因可能具有 3种不同的转录本。Northern杂交结果显示 ,此基因在所检测的 8种组织中除脑组织不表达外 ,其他 7种组织中均存在一种大小基本一致的转录本 ,约 2 .4kb ,该转录本在肾脏和骨骼肌中高度表达 ,在胎盘、心脏、肺脏中中度表达 ,在胰腺、肝脏中低度表达。杂交结果还显示 ,此基因在胰腺和胎盘组织中存在一种约 3.5kb的条带 ,在骨骼肌中存在另一种约 5 .0kb的条带。结论 电子杂交步移预测及Northern杂交结果证实了HumCyr6 1基因变异剪接位点的存在。
Objective To predict the mutated spliced transcripts of the HumCyr6 1 gene and to verify its expression profile in different tissues for preliminary investigation of the function of the HumCyr6 1 gene. Methods Using a computer-assisted electronic hybridization walking strategy, the mutated spliced transcripts of the HumCyr6 1 gene were predicted by comparing the ESTs returned from the query database and the previously cloned HumCyr6 1 cDNA sequence. The HumCyr6 1 gene fragment was prepared and hybridized with an MTNI membrane containing human tissue mRNA to perform expression profiling. RESULTS: The computer predicted three potential splice sites for the HumCyr6 1 gene, suggesting that the gene may have three different transcripts. Northern blotting results showed that except for no expression in the brain tissues of the 8 tissues tested, there was a transcript of approximately the same size in the other 7 tissues, approximately 2-4 kb. The transcript was found in the kidneys and bones. It is highly expressed in muscle, moderately expressed in the placenta, heart, and lung, and expressed low in the pancreas and liver. The hybridization results also showed that there is a band of about 3.5 kb in the pancreas and placenta, and another band of about 5.0 kb exists in the skeletal muscle. Conclusion The results of electronic hybridization walking prediction and Northern blotting confirmed the existence of variant splice sites of HumCyr6 1 gene.