黄脂菌素生物合成基因簇中调控基因xanR3的功能
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国家自然科学基金(31630002,31170085,31470183)


Function of AsrR family transcriptional regulatory gene xanR3 in the biosynthetic cluster of xantholipin
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    [目的]研究黄脂菌素产生菌灰黄链霉菌中编码ArsR家族转录调控蛋白(Arsenical resistance regulator)的xanR3基因的功能。[方法]利用大肠杆菌和链霉菌双亲本接合转移的方法,构建xanR3基因缺失突变株及回补突变株。利用cDNA在相邻同方向的基因间隔区进行PCR确定黄脂菌素生物合成基因簇中的转录单元。利用荧光定量RT-PCR方法进行突变株中黄脂菌素生物合成基因簇转录水平的检测。[结果]对得到的xanR3基因缺失突变株及回补突变株进行发酵,发现xanR3基因缺失突变株产黄脂菌素能力下降,回补菌株中黄脂菌素产量相比缺失突变株有一定程度的恢复,但仍未达到野生型水平。经鉴定,黄脂菌素生物合成基因簇中共有18个共转录单元,其中4个共转录单元在∆xanR3突变株中转录水平明显下降。[结论]ArsR家族转录调控基因xanR3是黄脂菌素生物合成的正调控基因。

    Abstract:

    [Objective] We revealed the function of the ArsR (Arsenical resistance regulator) family regulator XanR3 in xantholipin biosynthetic gene cluster. [Methods] Genetically, the xanR3 disrupted mutant CGF03 and complementary strain CGF06 were constructed by conjugation between S. flavogriseus SIIA-A02191 and Escherichia coli ET12567/pUZ8002. The operons within xantholipin biosynthesis cluster were grouped by RT-PCR and the transcriptional level was analyzed in the mutant CGF-3 by quantitative real-time RT-PCR. [Results] Inactivation of the xanR3 gene dramatically reduced xantholipin biosynthesis, and its complementation partially restored xantholipin production. Totally, genes in xantholipin biosynthetic gene cluster were assigned into 18 co-transcription units, and the disruption of xanR3 directly down-regulated the transcription of four operons as xanO1-xanS1, xanB1-xanB3, xanT-xanO9 and xanO10-xanC3.[Conclusion] XanR3, a ArsR family transcriptional regulator, is an activator of xantholipin biosynthesis.

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陈国锋,孔令新,王辂,褚以文,由德林. 黄脂菌素生物合成基因簇中调控基因xanR3的功能[J]. 微生物学报, 2018, 58(3): 401-410

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  • 收稿日期:2017-04-14
  • 最后修改日期:2017-06-14
  • 在线发布日期: 2018-02-12
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