Nishiyama Norihito, Mikami Kazuyuki, Ochiai Takehiko, Yamauchi Kiyoshi
Department of Biological Science, Faculty of Science, Shizuoka University, Shizuoka, Japan.
Zoolog Sci. 2009 Apr;26(4):259-65. doi: 10.2108/zsj.26.259.
We detected an unexpected small-sized DNA fragment during polymerase chain reaction (PCR) analysis of the heterogeneity of a macronuclear intergenic region of Paramecium caudatum. Southern blotting of total genomic DNA with the PCR product as a probe indicated that the small-sized DNA fragment constituted part of the macronuclear genome. Sequencing revealed that the PCR product was a chimeric DNA structure that may be generated by tail-to-tail fusion of the 5' region of the hemoglobin (hb) gene to most of the nucleosome assembly protein-1 (nap-1) gene. Short tandem repeats consisting of tetra- and tri-nucleotides exist at the putative cleavage sites in the hb and nap-1 genes, respectively. This feature differs from those found at the boundaries of TA-internal eliminated sequences in the P. aurelia complex and at transposable elements in other species. This suggests that the chimeric DNA is generated by a novel mechanism. Although the chimeric DNA contains the hb and nap-1 promoters, transcripts corresponding to the chimeric DNA were not detected by reverse transcription (RT)-PCR analysis during vegetative cell growth. Possible roles of chimeric DNA are discussed.
在对尾草履虫大核基因间隔区的异质性进行聚合酶链反应(PCR)分析过程中,我们检测到一个意外的小尺寸DNA片段。以PCR产物为探针,对总基因组DNA进行Southern印迹分析表明,该小尺寸DNA片段构成了大核基因组的一部分。测序结果显示,PCR产物是一种嵌合DNA结构,可能是由血红蛋白(hb)基因的5'区域与大部分核小体组装蛋白-1(nap-1)基因进行尾对尾融合产生的。在hb基因和nap-1基因的假定切割位点分别存在由四核苷酸和三核苷酸组成的短串联重复序列。这一特征不同于在 aurelia 复合种中TA内部消除序列边界处以及其他物种转座元件处发现的特征。这表明这种嵌合DNA是通过一种新机制产生的。虽然嵌合DNA含有hb和nap-1启动子,但在营养细胞生长期间,通过逆转录(RT)-PCR分析未检测到与嵌合DNA对应的转录本。文中讨论了嵌合DNA可能的作用。