Cloning and Quantitative Analysis of Genes Specifically Expressed in Mycelium and Primordium of Pleurotus ostreatus

机译：平菇侧耳菌丝体和原基中特异表达基因的克隆与定量分析

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Digital gene expression profilings (DGE) of two important developmental stages of Pleurotus ostreatus, mycelium and primordium, were constructed using next generation sequencing technology. To clone the specifically expressed genes, we use NlaⅢ digested double-strand cDNA of P. ostreatus as template, unmatched tag as upstream primer, AUAP adaptor primer (5'-GGCCACGCGTCGACTAGAC-3') as downstream primer for PCR amplification to get corresponding 3'cDNA sequence. Totally 120 tags of high, medium, and low expression level in mycelium and primordium of P. ostreatus were selected. After sequencing and blast, we obtain 3' cDNA sequences of 42 genes, among which 18 genes expressed specifically in the mycelium stage and 24 genes are specifically expressed in primordium stage. Full length cDNA sequences of hydrophobin 2 and myosin regulatory light chain genes were obtained by 5' RACE, meanwhile their expression level were confirmed by real-time fluorescence quantitative PCR. This study has laid the foundation for further study molecular mechanism of differentiation and development from mycelium to primordium in P. ostreatus.

机译：利用下一代测序技术构建了平菇的两个重要发育阶段的数字基因表达谱（DGE），即菌丝体和原基。为了克隆特异表达的基因，我们用NlaⅢ消化的平菇双链cDNA作为模板，未匹配的标签作为上游引物，AUAP衔接子引物（5'-GGCCACGCGTCGACTAGAC-3'）作为下游引物进行PCR扩增，得到相应的3。 cDNA序列。总共选择了120个在P. ostreatus的菌丝体和原基中高，中和低表达水平的标签。经过测序和鉴定后，我们获得了42个基因的3'cDNA序列，其中18个基因在菌丝体阶段特异性表达，24个基因在原基期特异性表达。通过5'RACE获得了疏水蛋白2和肌球蛋白调节性轻链基因的全长cDNA序列，并通过实时荧光定量PCR确定了它们的表达水平。该研究为进一步研究平菇中菌丝体向原基分化和发育的分子机理奠定了基础。

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来源
《Proceedings of the 18th congress of the International Society for Mushroom Science》|2012年|161-167|共7页
会议地点 Beijing(CN)
作者
Chaomin Yin; Yeye Chen; Hanyu Zhu; Jinghang Lei; Aimin Ma;
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作者单位

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China;

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China;

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China;

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China;

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China;

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原文格式 PDF
正文语种 eng
中图分类菌类（食用菌）;
关键词
Pleurotus ostreatus; Specifically Expressed Gene; Mycelium; Primordium; DGE; RACE; qPCR;

机译：平菇;特异表达基因;菌丝体;原mor; DGE; RACE; qPCR;

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2. Isolation and characterization of differentially expressed genes in the mycelium and fruit body of Pleurotus ostreatus [J] . Joongho Joh, Young-Jin Park, Eun-Suk Son, African Journal of Biotechnology . 2013,第24期

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3. Cloning of glyceraldehyde-3-phosphate dehydrogenase genes from the basidiomycete mushroom Pleurotus ostreatus and analysis of their expression during fruit-body development. [J] . Tasaki Y., Sato R., Toyama S., Mycoscience . 2014,第4期

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4. Cloning and Quantitative Analysis of Genes Specifically Expressed in Mycelium and Primordium of Pleurotus ostreatus [C] . Chaomin Yin, Yeye Chen, Hanyu Zhu, Congress of the International Society for Mushroom Science . 2012

机译：在菌丝体和肺炎菌菌丝菌丝体和原基中特异性表达的基因的克隆和定量分析
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6. iTRAQ-Based Quantitative Proteomic Analysis Reveals Proteomic Changes in Mycelium of Pleurotus ostreatus in Response to Heat Stress and Subsequent Recovery [O] . Yajie Zou, Meijing Zhang, Jibin Qu, -1

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7. Serial analysis of gene expression during mycelium and primordium stages of shiitake mushroom (xianggu) Lentinula edodes. [O] . 2003

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Cloning and Quantitative Analysis of Genes Specifically Expressed in Mycelium and Primordium of Pleurotus ostreatus

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