An Appropriate Increase in the Transcription of Aspergillus nidulans uvsC Improved Gene Targeting Efficiency
An Appropriate Increase in the Transcription of Aspergillus nidulans uvsC Improved Gene Targeting Efficiency
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DOI：
10.1271/bbb.68.1649
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发表时间：
2004-01
期刊：
影响因子：
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通讯作者：
Toyoaki Natsume;M. Egusa;M. Kodama;Richard D. Johnson;T. Itoh;Y. Itoh
中科院分区：
文献类型：
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作者：
Toyoaki Natsume;M. Egusa;M. Kodama;Richard D. Johnson;T. Itoh;Y. Itoh
由于基因组学研究的最新进展，基因靶向敲除特定基因的活性变得非常重要。但由于遗传转化过程中异位整合的高背景，这种技术仍然无法用于许多生物，包括具有重要经济意义的微生物。提高靶向效率的策略包括操纵参与同源重组的基因的表达。在本研究中，我们分别利用假芽曲霉和稻曲霉的甘油醛-3-磷酸脱氢酶和taka -淀粉酶A基因的启动子序列，提高了假芽曲霉uvsC的转录。虽然在3个位点上观察到靶向效率提高了几倍，但在具有较高uvsC转录水平的菌株中，菌丝生长受到抑制。这些结果表明，uvsC是基因靶向的限速因子，并且这种靶向效率的提高受到转录增加对细胞增殖的负面影响的阻碍。
Gene targeting to knock out the activity of specific genes has become important due to recent progress in genomics research. But this technique is still unavailable for many organisms, including economically important microorganisms, due to the high background of ectopic integration during genetic transformation. Strategies to improve targeting efficiency have included manipulating the expression of genes that are involved in homologous recombination. In this study, transcription of Aspergillus nidulans uvsC was elevated using the promoter sequences of the glyceraldehyde-3-phosphate dehydrogenase and Taka-amylase A genes from A. nidulans and A. oryzea respectively. Although a several-fold increase in the efficiency of targeting was observed at 3 loci, mycelial growth was suppressed in strains that had higher levels of uvsC transcription. These results suggest that uvsC is a rate-limiting factor in gene targeting, and that the increased efficiency of this targeting is hindered by a negative effect of increased transcription on cell proliferation.