Transcriptome profiles for high-cell-density recombinant and wild-type Escherichia coli
Transcriptome profiles for high-cell-density recombinant and wild-type Escherichia coli
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DOI：
10.1002/bit.20340
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发表时间：
2005-04-20
影响因子：
3.8
通讯作者：
Harcum, SW
中科院分区：
文献类型：
--
作者：
Haddadin, FT;Harcum, SW
通过DNA微阵列分析了在良好控制的同步高细胞密度补料分批培养期间野生型（无质粒）和重组（携带质粒）大肠杆菌的转录组谱。据观察，生长阶段显着影响转录组图谱，并且重组和野生型培养物的转录组图谱显着不同。检查了野生型和重组培养物对添加异丙基-1-硫代-β-D-半乳糖-吡喃糖苷- (IPTG-) 的反应，其中 IPTG 在带有质粒的培养物中诱导重组蛋白的产生。添加 IPTG 显着改变了进入稳定期的野生型培养物的转录组反应。 IPTG诱导的重组蛋白产生导致许多能量合成基因（atp、nuo、cyo）以及几乎所有转录和翻译相关基因（rpo、rpl、rpm、rps、rrf、rrl、rrs）显着下调。由于 IPTG 诱导，许多噬菌体（psp、htl）和转座子相关基因（tra、ins）在重组培养物中受到显着调节。这些结果表明，与重组蛋白生产相关的信号传导机制可能会以噬菌体防御机制的形式诱导代谢负担。总而言之，这些结果表明重组蛋白的产生引发了一系列转录组反应，下调了维持生产力所需的基因。 (c) 2005 年 Wiley 期刊公司。
The transcriptome profiles for wild-type (plasmid-free) and recombinant (plasmid-bearing) Escherichia coli during well-controlled synchronized high-cell-density fed-batch cultures were analyzed by DNA microarrays. It was observed that the growth phase significantly affected the transcriptome profiles, and the transcriptome profiles were significantly different for the recombinant and wild-type cultures. The response of the wild-type and recombinant cultures to an isopropyl-1-thio-beta-D-galacto-pyranoside- (IPTG-) addition was examined, where IPTG induced recombinant protein production in the plasmid-bearing cultures. The IPTG-addition significantly altered the transcriptome response of the wild-type cultures entering the stationary phase. The IPTG-induced recombinant protein production resulted in a significant down-regulation of many energy synthesis genes (atp, nuo, cyo), as well as nearly all transcription- and translation-related genes (rpo, rpl, rpm, rps, rrf, rrl, rrs). Numerous phage (psp, htl) and transposon-related genes (tra, ins) were significantly regulated in the recombinant cultures due to the IPTG-induction. These results indicate that the signaling mechanism, associated with the recombinant protein production, may induce a metabolic burden in the form of a phage defense mechanism. Taken together, these results indicated that recombinant protein production initiated a cascade of transcriptome responses that down-regulated the very genes needed to sustain productivity. (c) 2005 Wiley Periodicals, Inc.