Use of segment-based microarray in the analysis of global gene expression in response to various environmental stresses in the cyanobacterium Anabaena sp PCC 7120
Use of segment-based microarray in the analysis of global gene expression in response to various environmental stresses in the cyanobacterium Anabaena sp PCC 7120
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DOI：
10.2323/jgam.50.1
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发表时间：
2004-02-01
影响因子：
1.2
通讯作者：
Kuhara, S
中科院分区：
文献类型：
--
作者：
Sato, N;Ohmori, M;Kuhara, S
我们制备了包含形成异胞的丝状蓝藻Anabaena sp.基因组序列的微阵列。PCC7120。该蓝藻的全基因组编码约5,368个蛋白质编码基因，位于6.4MbP的主染色体上。从测序克隆中筛选出2,407个DNA片段，经聚合酶链式反应扩增后，在玻片上一式两份进行斑点。这些微阵列与广泛使用的商业或定制的其他微生物微阵列的不同之处在于，每个DNA片段长3-4kbp，平均包含约3-4个预测基因。然而，这一特征并没有降低微阵列的实用性，因为我们能够检测到一些潜在的新基因，这些基因是在氮缺乏、低温和干旱的情况下诱导产生的。此外，我们还发现了一些基因组区域，在这些区域中数十个相邻的基因同时受到调控。这些结果表明，这些基于片段的微阵列特别适用于像鱼腥藻这样的大基因组，其基因数量超过了技术或实际限制。
We prepared microarrays that contain genomic sequences of a heterocyst-forming filamentous cyanobacterium Anabaena sp. PCC 7120. The complete genome of this cyanobacterium codes for about 5,368 protein-coding genes in the main chromosome of 6.4 Mbp. In total, 2,407 DNA segments were selected from the sequencing clones, and amplified by PCR, then spotted on glass slides in duplicate. These microarrays differ from the widely used commercial or custom-made ones for other microorganisms in that each DNA segment was 3-4 kbp long, and contained about 3-4 predicted genes on average. This feature, however, did not decrease the usefulness of the microarrays, since we were able to detect a number of potentially novel genes that are induced in response to nitrogen deprivation, low temperature and drought. In addition, we found some genomic regions in which dozens of contiguous genes are simultaneously regulated. These results suggest that these segment-based microarrays are useful especially for such large genomes as Anabaena, for which the number of genes exceeds either technical or practical limitations.