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利用RAPD差异显示技术分离糙皮侧耳子实体发育过程中差异表达的基因。

Isolation of genes differentially expressed during the fruit body development of Pleurotus ostreatus by differential display of RAPD.

作者信息

Sunagawa Masahide, Magae Yumi

机构信息

Department of Applied Microbiology, Forestry and Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan.

出版信息

FEMS Microbiol Lett. 2005 May 15;246(2):279-84. doi: 10.1016/j.femsle.2005.04.018.

Abstract

To analyze genes involved in fruit body development of Pleurotus ostreatus, mRNAs from three different developmental stages: i.e., vegetative mycelium, primordium, and mature fruit body, were isolated and reverse-transcribed to cDNAs. One hundred and twenty random PCR amplifications were performed with the cDNAs, which generated 382, 394, 393 cDNA fragments from each developmental stage. From these fragments, four cDNA clones specifically expressed in primordium or mature fruit body were detected. Sequence analysis and database searches revealed significant similarity with triacylglycerol lipase, cytochrome P450 sterol 14 alpha-demethylase and developmentally regulated genes of other fungi. Northern blot analyses confirmed that all of the four cDNAs were unexpressed in mycelium, thus stage-specific genes for fruit body formation of P. ostreatus were successfully isolated.

摘要

为分析参与平菇子实体发育的基因,从三个不同发育阶段(即营养菌丝体、原基和成熟子实体)分离mRNA,并反转录成cDNA。用这些cDNA进行了120次随机PCR扩增,每个发育阶段分别产生了382、394和393个cDNA片段。从这些片段中,检测到四个在原基或成熟子实体中特异性表达的cDNA克隆。序列分析和数据库检索显示,它们与三酰基甘油脂肪酶、细胞色素P450固醇14α-脱甲基酶以及其他真菌的发育调控基因具有显著相似性。Northern印迹分析证实,这四个cDNA在菌丝体中均未表达,因此成功分离出了平菇子实体形成的阶段特异性基因。

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