Expression profiling by whole-genome interspecies microarray hybridization reveals differential gene expression in procyclic promastigotes, lesion-derived amastigotes, and axenic amastigotes in Leishmania mexicana
Expression profiling by whole-genome interspecies microarray hybridization reveals differential gene expression in procyclic promastigotes, lesion-derived amastigotes, and axenic amastigotes in Leishmania mexicana
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DOI：
10.1016/j.molbiopara.2005.12.009
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发表时间：
2006-04-01
影响因子：
1.5
通讯作者：
Forney, JD
中科院分区：
文献类型：
--
作者：
Holzer, TR;McMaster, WR;Forney, JD
我们研究了墨西哥利什曼原虫的转录组，使用高密度全基因组寡核苷酸微阵列来识别差异调节的mrna，这些芯片是由一个密切相关的物种利什曼原虫的基因组数据设计的。对8156个预测编码区的阵列杂交数据进行统计分析，发现288个基因（占所有基因的3.5%）的稳态mRNA水平符合原无性体和病变源性无性体的差异调控标准。有趣的是，在原生无尾线虫和雌雄无尾线虫的样本比较中，只有17个基因（0.2%）符合相同的差异调控统计标准。受调控基因数量的减少是在无菌条件下细胞中转录水平增加的结果。通过定量PCR验证了一部分基因的表达数据。我们的研究表明，微阵列上的种间杂交可以用于分析密切相关的原生动物寄生虫，无性系培养条件可能会改变无性系培养的转录物丰度，并且在形态不同的promastigote和amastigote培养的细胞之间，少数mrna的丰度只有相对适度的变化。利什曼原虫可能代表了真核生物分化的另一种范式，mRNA丰度变化的贡献最小。(c) 2005 Elsevier B.V.版权所有
We examined the Leishmania mexicana transcriptome to identify differentially regulated mRNAs using high-density whole-genome oligonucleotide microarrays designed from the genome data of a closely related species, Leishmania major. Statistical analysis on array hybridization data representing 8156 predicted coding regions revealed 288 genes (3.5% of all genes) whose steady-state mRNA levels meet criteria for differential regulation between promastigotes and lesion-derived amastigotes. Interestingly, sample comparison of promastigotes to axenic amastigotes resulted in only 17 genes (0.2%) that meet the same statistical criteria for differential regulation. The reduced number of regulated genes is a consequence of an increase in the magnitude of the transcript levels in cells under axenic conditions. The expression data for a subset of genes was validated by quantitative PCR. Our studies show that interspecies hybridization on microarrays can be used to analyze closely related protozoan parasites, that axenic culture conditions may alter amastigote transcript abundance, and that there is only a relatively modest change in abundance of a few mRNAs between morphologically distinct promastigote and amastigote cultured cells. Leishmania may represent an alternative paradigm for eukaryotic differentiation with minimal contributions from changes in mRNA abundance. (c) 2005 Elsevier B.V. All rights reserved.