人类Cyr61基因的组织表达谱分析

Studies on the Tissue Expression Pattern of HumCyr61

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 study the function of HumCyr61, three alternative splicing transcripts were speculated and confirmed afterwards.Methods With the help of computer, electronic hybridization walking strategy was used to speculate alternative splicing of HumCyr61 by comparison of the BLAST result obtained from dbEST and HumCyr61 cDNA sequence. Northern blotting was performed to hybridize α- 32 P-dATP labeled HumCyr61 cDNA partial sequence with mRNA of 8 different human tissues that had been transferred to the MTNI membrane.Results Northern hybridization showed that HumCyr61 was expressed as a～2.4kb transcript in almost all tissues tested except brain, among which high expression in kidney and skeletal muscle, medium expression in placenta, heart, lung, and low expression in pancreas and liver. Northern hybridization also showed that there were another transcript of～3.5kb in pancreas, placenta and transcript of～5.0kb in skeletal muscle. The result is consistent with the previous speculation obtained with computer.Conclusion There exist two potential transcripts except the cloned HumCyr61 cDNA.