Isolation of thecandidate gene for the marker of the body time determinant in human saliva.

利用人唾液腺细胞系HSG，通过芯片上芯片（ChIP on ChIP）和微阵列（microarray）相结合的实验，鉴定出了22个由时钟基因直接控制并表现出昼夜节律转录的基因。22个基因中含有Rev-erb α作为阳性对照。然后，通过RT-PCR选择Rev-erb α等9个基因作为正常人唾液腺中表达的基因。在这9个基因中，除Rev-erb α外，我们发现3个基因具有明显的昼夜节律转录。最后通过western blot分析成功鉴定了唾液中该基因的产物。这些结果表明，所选择的基因是人唾液中身体时间决定因素标记的良好候选基因。

We have identified 22 genes which are controlled directly by clock genes and show circadian transcription by the combination of ChIP on Chip and microarray experiments using human salivary gland cell line HSG. Rev-erb alpha was included in the 22 genes as a positive control. Then, 9 genes including Rev-erb alpha were selected as genes expressing in the normal human salivary gland by RT-PCR. In these 9 genes we found that 3 genes except Rev-erb alpha showed clear circadian transcription. Finally we have succeeded to identify the gene whose product is detected in saliva by western blot analysis. These results suggest that the selected gene is the good candidate for the marker of the body time determinant in human saliva.