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微小泰勒虫两个单拷贝核糖体转录单元内LSU rRNA基因的微序列异质性与表达

Microsequence heterogeneity and expression of the LSU rRNA genes within the two single copy ribosomal transcription units of Theileria parva.

作者信息

Bishop R, Gobright E, Spooner P, Allsopp B, Sohanpal B, Collins N

机构信息

International Livestock Research Institute (ILRI), P.O. Box 30709, Nairobi, Kenya.

出版信息

Gene. 2000 Oct 31;257(2):299-305. doi: 10.1016/s0378-1119(00)00386-3.

Abstract

The nucleotide sequences of the large subunit ribosomal RNA coding genes within the two single copy ribosomal DNA transcription units of a cloned Theileria parva isolate from a buffalo were determined. The two LSU rRNA coding units differed by 11 nucleotide substitutions and two deletions of 1 and 6 bp, all located in the 5' end of the LSU coding region. We also observed microsequence heterogeneity between the two buffalo parasite LSU sequences and the previously determined LSU rRNA gene of a T. parva parasite isolated from cattle. At all positions which were variable between the two LSU rRNA coding sequences of the buffalo-derived parasite, either unit 1 or unit 2 matched the LSU rRNA coding sequence of the cattle-derived T. parva parasite in a mosaic pattern. Synthetic oligonucleotides specific for LSU units 1 and 2 of the buffalo T. parva were developed, and used to assay expression of the two units. Both units were expressed in the intra-lymphocytic schizont stage of T. parva. A 2.5-10-fold excess of rRNA derived from LSU unit 1, compared with unit 2, was observed in the schizont stage, the difference being attributable to variation in the level of expression of unit 2. Theileria represents the third genus of Sporozoan protozoa, in addition to Plasmodium and Babesia, exhibiting rRNA coding genes, which are divergent in sequence between different transcription units.

摘要

测定了从一头水牛身上克隆的泰勒虫分离株的两个单拷贝核糖体DNA转录单元内的大亚基核糖体RNA编码基因的核苷酸序列。这两个大亚基rRNA编码单元有11个核苷酸替换和两个1bp和6bp的缺失,均位于大亚基编码区的5'端。我们还观察到这两个水牛寄生虫大亚基序列与先前测定的从牛分离的泰勒虫寄生虫的大亚基rRNA基因之间存在微序列异质性。在水牛源寄生虫的两个大亚基rRNA编码序列之间可变的所有位置上,单元1或单元2以镶嵌模式与牛源泰勒虫寄生虫的大亚基rRNA编码序列匹配。开发了针对水牛泰勒虫大亚基单元1和单元2的合成寡核苷酸,并用于检测这两个单元的表达。两个单元均在泰勒虫的淋巴细胞内裂殖体阶段表达。在裂殖体阶段观察到,与单元2相比,来自大亚基单元1的rRNA过量2.5至10倍,差异归因于单元2表达水平的变化。泰勒虫是除疟原虫和巴贝斯虫之外的第三个孢子虫原生动物属,其表现出在不同转录单元之间序列不同的rRNA编码基因。

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