Iiyama Kazuhiro, Chieda Yuuka, Yasunaga-Aoki Chisa, Hayasaka Shoji, Shimizu Susumu
Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka 812-8581, Japan.
J Eukaryot Microbiol. 2004 Nov-Dec;51(6):598-604. doi: 10.1111/j.1550-7408.2004.tb00592.x.
Ribosomal DNA (rDNA) containing small subunit (SSU) rDNA and both flanking regions in the entomopathogenic microsporidian Nosema bombycis NIS 001 was amplified from genomic DNA with a primer set based on the sequence of an inverse polymerase chain reaction (PCR)-derived fragment. In this fragment, SSU rDNA was divided by a 618-bp insert at nt 599, and 5S rDNA was located downstream of the SSU rDNA, fragmented by 284-bp intergenic spacer. In addition, the 48-bp 3'-end of large subunit (LSU) rDNA was located 118 bp upstream of the fragmented SSU rDNA. In the amplicon, the region upstream of the LSU rDNA was a homologue of the C-terminal CHARLIE8 transposon-like element of human GTF2IRD2. In this organism, another fragmented SSU rDNA, which was divided by a 231-bp insert at nt 50, was also detected. Both the intact (insertless) and fragmented SSU rDNAs clustered with LSU rDNA and 5S rDNA and the intergenic sequences between SSU rDNA and 5S rDNA were divergent in an organism. Reverse transcription (RT)-PCR assay indicated that not only the intact SSU rDNA but also the fragmened SSU rDNA were transcribed in N. bombycis.
从昆虫病原微孢子虫家蚕微孢子虫NIS 001的基因组DNA中,使用基于反向聚合酶链反应(PCR)衍生片段序列的引物对,扩增出包含小亚基(SSU)核糖体DNA(rDNA)及其两侧区域的核糖体DNA(rDNA)。在这个片段中,SSU rDNA在第599位核苷酸处被一个618 bp的插入片段隔开,5S rDNA位于SSU rDNA的下游,被284 bp的基因间隔区隔开。此外,大亚基(LSU)rDNA的48 bp 3'末端位于断裂的SSU rDNA上游118 bp处。在扩增子中,LSU rDNA上游区域与人GTF2IRD2的C末端CHARLIE8转座子样元件同源。在这种生物中,还检测到另一个断裂的SSU rDNA,它在第50位核苷酸处被一个231 bp的插入片段隔开。完整(无插入)和断裂的SSU rDNA都与LSU rDNA和5S rDNA聚类,并且在一种生物中,SSU rDNA和5S rDNA之间的基因间隔序列是不同的。逆转录(RT)-PCR分析表明,完整的SSU rDNA和断裂的SSU rDNA在家蚕微孢子虫中都被转录。