Molecular mechanisms of circadian rhythm system and sleep disorders

1)我们克隆了人BMAL 1基因的BAC克隆，并测定了其序列。结果表明，人BMAL 1基因编码区由17个外显子组成，有几个BMAL 1变异体是剪接变异体的产物。2）克隆了小鼠BMAL 1的BAC克隆。3）测定小鼠BMAL 1启动子区序列。我们确定了BMAL 1的启动子/增强子区域。4）确定了BMAL 1的转录起始位点。5）搜索了睡眠障碍患者BMAL 1基因的多态性。在BMAL 1编码区未发现多态性; 6）建立了检测clock基因产物相互作用的双杂交系统; 7）建立了检测mPer 1启动子激活能力的荧光素酶检测系统; 8）制备了clock基因融合蛋白，用于检测clock基因产物相互作用; 9）克隆了一个bHLH-PAS因子，命名为BMAL2.10）检测BMAL 2的表达、染色体定位和细胞定位。

1) We have cloned a BAC clone for human BMAL1 gene and determined its sequence. It revealed that human BMAL1 coding region consists from 17 exons and several BMAL1 variants are products from splicing variants.2) We have cloned a BAC clone for mouse BMAL1. Its coding region splits over 30kb.3) Mouse BMAL1 promoter region was sequenced. We identified the promoter/enhancer region of BMAL1.4) The transcription start sites for BMAL1 have been identified.5) Polymorphism of BMAL1 gene from sleep disorders were searched. No polymorphism were identified in the BMAL1 coding region.6) Two-hybrid system to test the interaction of clock gene products have been established.7) Luciferase assay system to test the ability to activate mPer1 promoter have been established.8) Fusion proteins of clock gene have been prepared for examining the interaction of clock gene products.9) A nobel bHLH-PAS factor have been cloned and designated BMAL2.10) The expression, chromosomal localization and cellular localization of BMAL2 were examined.