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为了研究冠突散囊菌nsdD基因的功能,本研究将冠突散囊菌nsdD的cDNA置于强启动子三磷酸甘油醛脱氢酶启动子PgpdA下游及色氨酸合成酶C基因终止子TtrpC上游构建nsdD基因超表达载体PURT5750,采用根癌农杆菌介导方法将表达质粒PURT5750转化到冠突散囊菌进行功能验证。结果显示向基因组随机插入外源质粒DNA,随机挑选11个nsd D基因超表达转化子,对nsdD基因超表达转化子菌株培养观察并与野生型菌株相比发现,在5%和17%NaCl培养基中菌落差异不显著,而且仍能产生成熟的有性孢子。本实验为冠突散囊菌有性发育调控机制的研究提供了技术基础。
Abstract：To investigate the function of nsd D gene in Eurotium cristatum, the cDNA of nsdD gene was inserted into the downstream of promoter Pgpd A of glyceraldehyde 3 phosphoric acid dehydrogenase gene promoter and the upstream of terminator TtrpC of tryptophan synthetase to constructe the over-expression vector PURT5750 of nsdD gene, and the PURT5750 expression plasmid was transformed into Eurotium cristatum by Agrobacterium-mediated transformation to verify its functions. The result showed that exogenous plasmid DNA was randomly inserted into the genome, and eventually 11 nsdD gene over-expression transformants were selected. The nsdD gene over-expression transformant strain was cultured, observed and compared with wildtype strain, of which the result showed that there was no difference in the 5% and 17% NaCl medium, and it still formed mature sexual spores. The work provided a technical basisfor the future reseach ofmechanism ofsexual development of Eurotium cristatum.
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Zhu L.L.,Tan Y.M.,Ge Y.Y.,Wang Y.C.,Liu Y.X.,and Liu Z.Y.2016,Detection of“Golden Flower”(Eurotium cristatum)in the Producing process of fuzhuan Tea,Jiyinzuxue Yu Yingy ong Shengwuxue(Genomics and Applied Biology),35(1)124-129(朱李丽,谭玉梅,葛永怡,王玉臣,刘永翔,刘作易2016,茯砖茶生产过程中“金花”菌的检测,基因组学与应用生物学,35(1):124-129)
基本信息:
DOI：10.13417/j.gab.036.000900
中图分类号:Q78;Q93
引用信息:
[1]余春芳,熊庆,李文仿,等.冠突散囊菌nsdD基因超表达菌株的构建及表型分析[J].基因组学与应用生物学,2017,36(03):900-905.DOI:10.13417/j.gab.036.000900.
基金信息:
十堰市科技局项目(ZD2012019);; 湖北医药学院校基金(2014QDJZR15)共同资助
2016-10-24
2016-10-24
2016-10-24