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利用抑制性差减杂交技术鉴定谢瓦氏曲霉间型变种产孢相关的基因

利用抑制性差减杂交技术鉴定谢瓦氏曲霉间型变种产孢相关的基因

ISSN:1672-6472
2013年第32卷第1期
研究论文
谭玉梅[1]    王海[1]    刘永翔[2]    刘作易[3] TAN Yu-Mei,WANG Hai,LIU Yong-Xiang and LIU Zuo-Yi
谭玉梅 (华中农业大学生命科学技术学院 湖北武汉430000;贵州省生物技术研究所 贵州贵阳550006); 王海 (华中农业大学生命科学技术学院 湖北武汉430000;贵州省生物技术研究所 贵州贵阳550006); 刘永翔 (贵州省生物技术研究所 贵州贵阳550006); 刘作易(贵州省农业科学院 贵州贵阳550006); TAN Yu-Mei 1, 2 WANG Hai 1, 2 LIU Yong-Xiang 2 LIU Zuo-Yi 3 1 College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430000, China 2 Guizhou Institute of Biotechnology, Guiyang, Guizhou 550006, China 3 Guizhou Academy of Agricultural Science, Guiyang, Guizhou 550006, China

谢瓦氏曲霉间型变种Aspergillus chevalieri var.intermedius俗称"金花菌",是茯砖茶发酵中的优势菌种,该菌在不同的渗透压胁迫下产生不同类型的孢子。采用抑制性差减杂交技术(suppression subtractive hybridization,SSH),以低渗条件下产生的子囊孢子和高渗条件下产生的分生孢子为材料分别与营养菌丝体构建两个正反向文库。每个库随机挑选400个阳性克隆进行测序。然后将所得到的基因片段在GenBank进行BLASTx和BLASTn同源比对,筛选出的差异表达的基因涉及蛋白质代谢、细胞代谢、转录调控等多个方面,其中可能参与有性产孢调控的基因有13条,可能参与无性产孢调控的基因有10条。

Aspergillus chevalieri var. intermedius was the dominant fermentative fungus in ‘fuzhuan’ tea. This fungus formed different kinds of spores under different osmotic pressure. The ascospore and conidia produced under low and high osmotic pressure, respectively, were used to construct suppression subtractive hybridization library with the vegetative mycelia as the control. Four hundred clones were selected from each library for nucleotide sequencing and the obtained sequences were searched against the GenBank database using the BLASTn and BLASTx programs. The differentiallyexpressed genes are related to protein metabolism, cell metabolism and transcriptional regulation, and so on. Among them, thirteen genes possibly related to sexual sporulation and ten genes possibly related to asexual sporulation were found.

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ISSN:1672-6472
2013年第32卷第1期
研究论文

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