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利用抑制差减杂交技术在灵芝液体静置培养的无性孢子形成过程中分离和分析差异表达基因。

Isolation and analysis of differentially expressed genes during asexual sporulation in liquid static culture of Ganoderma lucidum by suppression subtractive hybridization.

机构信息

State Key Laboratory of Bioreactor Engineering, School of Biotechnology, [corrected] East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.

出版信息

Mol Biol Rep. 2012 Apr;39(4):3603-10. doi: 10.1007/s11033-011-1134-2. Epub 2011 Jul 3.

Abstract

Ganoderma lucidum differentiates in liquid static culture by forming aerial mycelia and asexual spores, and this differentiation process is accompanied by higher production of anti-tumor compounds ganoderic acids. To gain an insight into the molecular events during asexual sporulation of G. lucidum, comparative transcriptome analysis using suppression subtractive hybridization (SSH) technique was performed to identify preferentially expressed genes in liquid static culture vs. in traditional shaking culture. After macroarray analysis of 1920 cDNAs from SSH library, 147 unigenes which exhibited high expression in static culture were identified. Among these sequences, putative translations of 88 unigenes possessed much similarity to known proteins involved in cell organization, signal transduction, cell metabolism, protein biosynthesis and transcription regulation; 13 had significant similarity to hypothetical proteins; the remaining 46 showed little or no similarity to GenBank sequences. RT-qPCR analysis confirmed increases in transcripts of selected genes under liquid static culture condition. The results of this study present the useful application of EST analysis on G. lucidum and provide preliminary indication of gene expression putatively involved in asexual sporulation process.

摘要

灵芝在液体静置培养中通过形成气生菌丝和无性孢子来分化,这个分化过程伴随着抗肿瘤化合物灵芝酸的更高产量。为了深入了解灵芝无性孢子形成过程中的分子事件,采用抑制性消减杂交(SSH)技术进行了比较转录组分析,以鉴定液体静置培养与传统摇瓶培养中优先表达的基因。对 SSH 文库的 1920 个 cDNA 进行宏阵列分析后,鉴定出了 147 个在静置培养中高表达的基因。在这些序列中,88 个推定翻译的基因与参与细胞组织、信号转导、细胞代谢、蛋白质生物合成和转录调节的已知蛋白质具有高度相似性;13 个与假定蛋白质具有显著相似性;其余 46 个与 GenBank 序列几乎没有相似性或没有相似性。RT-qPCR 分析证实了在液体静置培养条件下选定基因的转录物增加。这项研究的结果展示了 EST 分析在灵芝上的有用应用,并初步表明了基因表达可能参与无性孢子形成过程。

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