Yamamoto H, Naruse A, Ohsaki T, Sekiguchi J
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano.
J Biochem. 1995 Apr;117(4):888-96. doi: 10.1093/oxfordjournals.jbchem.a124792.
The nucleotide sequence of a large rRNA gene and its flanking regions in cloned fragments of mitochondrial DNA from a patulin producer, Penicillium urticae NRRL2159A, was determined by dideoxy sequencing, and the 5' end and intron-exon border of the 1-rRNA gene were determined by primer extension analysis and RNA sequencing, respectively. In addition to the extensive sequence homology of the 3' end of the P. urticae mt 1-rRNA gene with those of Aspergillus nidulans and Neurospora crassa, the P. urticae gene had a 1,685 bp intron which separates a 3,307 bp 5' exon and a 583 bp 3' exon. In spite of being closely related Penicillium species, the size of the 5' exon of the P. urticae mt 1-rRNA is 472 bp larger than that of P. chrysogenum, whereas the sizes of the 3' exon and intron of P. urticae are very similar to those of P. chrysogenum (581 bp for the 3' exon and 1,678 bp for the intron). The intron of P. urticae contains a structure similar to the consensus one of the self splicing group IA intron and a large open reading frame suggested to be a gene for ribosomal protein S5. A sequence similar to the I-SceI recognition sequence was found at the exon-intron border. Extensive sequence homology was observed between P. urticae and P. chrysogenum, exceptions being in four regions in the 5' exon. These non-homologous regions were located in the hairpin and variable regions outside of the core structures. Comparison of the mt 1-rRNA sequences of several filamentous fungi revealed that the above four non-homologous regions are greatly expanded, and two other non-homologous regions appear at the 3' ends of the 5' exon and 3' exon.
采用双脱氧测序法测定了棒曲霉素产生菌荨麻青霉NRRL2159A线粒体DNA克隆片段中一个大rRNA基因及其侧翼区域的核苷酸序列,并用引物延伸分析和RNA测序分别确定了1-rRNA基因的5'端和内含子-外显子边界。除了荨麻青霉mt 1-rRNA基因3'端与构巢曲霉和粗糙脉孢菌的3'端具有广泛的序列同源性外,荨麻青霉基因还有一个1685 bp的内含子,该内含子将一个3307 bp的5'外显子和一个583 bp的3'外显子分隔开。尽管荨麻青霉和产黄青霉是亲缘关系密切的青霉属物种,但荨麻青霉mt 1-rRNA的5'外显子大小比产黄青霉的大472 bp,而荨麻青霉3'外显子和内含子的大小与产黄青霉的非常相似(3'外显子为581 bp,内含子为1678 bp)。荨麻青霉的内含子包含一个与自我剪接IA组内含子的共有结构相似的结构以及一个推测为核糖体蛋白S5基因的大开放阅读框。在外显子-内含子边界发现了一个与I-SceI识别序列相似的序列。在荨麻青霉和产黄青霉之间观察到广泛的序列同源性,5'外显子中的四个区域除外。这些非同源区域位于核心结构之外的发夹和可变区域。几种丝状真菌的mt 1-rRNA序列比较表明,上述四个非同源区域大大扩展,另外两个非同源区域出现在5'外显子和3'外显子的3'端。