Ossorio P N, Sibley L D, Boothroyd J C
Department of Microbiology and Immunology, Stanford University School of Medicine, CA 94305-5402.
J Mol Biol. 1991 Dec 5;222(3):525-36. doi: 10.1016/0022-2836(91)90494-q.
In the course of our genetic studies on Toxoplasma gondii, it was discovered that one cosmid hybridized to a repetitive element. The hybridization pattern observed for the enzyme BglII indicated that this cosmid hybridized to a large number of discrete, but related elements. Four BglII fragments were subcloned from the cosmid, and each was shown to hybridize with all the others, as well as to numerous dispersed sequences in genomic DNA. Three subclones were sequenced in their entirety, and shown to contain fragments of the genes for cytochrome oxidase subunit I and apocytochrome b, complete and functional copies of which have been found in only mitochondrial genomes. All the subcloned fragments were bounded at both ends by a 91 base-pair sequence, which contains a site for BglII. This 91 base-pair sequence could be found as either a direct or inverted repeat. It was determined that the BglII elements are arrayed downstream from a single copy nuclear gene. Comparison of genomic and cosmid DNAs confirmed that the cosmid faithfully reflects the nuclear genome. Although the mitochondrial genome of Toxoplasma has not been characterized, these nuclear mitochondrial-like sequences appear to be internally rearranged with respect to known, functional mitochondrial genomes, and with respect to each other. The finding of short repeated sequences flanking these elements may be a clue to the mechanism of their dissemination.
在我们对刚地弓形虫的基因研究过程中,发现一个黏粒与一个重复元件杂交。用BglII酶观察到的杂交模式表明,这个黏粒与大量离散但相关的元件杂交。从该黏粒中克隆出四个BglII片段,结果显示每个片段都能与其他片段杂交,也能与基因组DNA中的许多分散序列杂交。对三个亚克隆进行了全序列测定,结果显示它们含有细胞色素氧化酶亚基I和脱辅基细胞色素b的基因片段,而完整且有功能的这些基因仅在线粒体基因组中发现过。所有亚克隆片段两端都由一个91个碱基对的序列界定,该序列含有一个BglII酶切位点。这个91个碱基对的序列可以以正向重复或反向重复的形式存在。已确定BglII元件排列在一个单拷贝核基因的下游。基因组DNA和黏粒DNA的比较证实,该黏粒忠实地反映了核基因组。虽然刚地弓形虫的线粒体基因组尚未被鉴定,但这些类核线粒体序列在内部似乎相对于已知的、有功能的线粒体基因组以及它们彼此之间发生了重排。在这些元件两侧发现短重复序列可能是其传播机制的一个线索。