球毛壳甲素（Chaetoglobosin A，ChA）是丝状真菌球毛壳菌的标志性代谢产物，具有强烈的抗肿瘤、杀线虫以及抑制植物病原真菌活性。因此，ChA具有开发为抗肿瘤药物或生物农药的潜在价值。目前，ChA生物合成基因簇已经明确，其调控机理还有待进一步研究。课题组前期筛选到一个锌指结构转录因子CgTF6，通过靶向敲除实验证明了CgTF6对球毛壳甲素合成的负反馈调节作用。此外还发现CgTF6不仅参与调控球毛壳甲素的生物合成，CgTF6功能缺失对球毛壳菌全基因组27个聚酮合酶基因表达也产生显著影响。为进一步确定CgTF6的细胞功能，对其进行亚细胞定位，并检测表观遗传修饰药物处理下CgTF6表达情况。本论文利用CRISPR-Cas9基因敲除体系，以△cgtf6为受体菌，以乳清酸核苷-5’-磷酸脱羧酶基因Cgura3为筛选标记，敲除Cgura3基因的同时回补yeGFP标记的Cgtf6基因。分别用自身启动子Ptf6驱动和强启动子Cgura3启动子驱动yeGFP-CgTF6融合蛋白表达，进行CgTF6亚细胞定位，进一步探究CgTF6的表达情况。通过无缝连接、TA克隆及重叠PCR等多种分子生物学手段获得带yeGFP标记的自启动子融合片段、强启动子融合片段及Cas9表达载体。利用qRT-PCR技术分析表观遗传修饰药物对野生型菌株Cgtf6转录水平的影响，结果显示组蛋白脱乙酰基酶抑制剂SAHA（Suberoylanilide hydroxamic acid）抑制Cgtf6转录，mRNA水平下调至野生型的0.3倍。本论文结果为CgTF6的功能研究提供一些载体及实验数据。

The representative natural product of the filamentous fungus Chaetomium globosum, is chaetoglobosin A (ChA) that has a variety of biological activities such as anti-tumor,nematicidal activities and significant growth inhibition effects on plant pathogenic fungi.It is suggested that ChA has the great potential to act as anti-tumor drugs or biological pesticides.Up to now, the biosynthesis gene cluster of ChA has been revealed,but the the regulatory mechanism of ChA biosynthesis is still unclear.Our team previously screened a putative zinc finger transcription factor CgTF6,which has been demonstrated to be acted as a negative feedback regulator on ChA biosynthesis by targeted gene deletion.In addition,CgTF6 also has a significant effect on the transcription of another 27 polyketide synthase in the not only involved in the regulation of ChA biosynthesis, but also has a significant effect on the expression of 27 polyketide synthase genes in the whole genome of Chaetomium globosum.For further verification of the function of CgTF6, we plan to observe the subcellular localization of CgTF6 and determin the expression pattern of CgTF6 in the presence of epigenetic modification drugs.In this paper, Cgtf6 deletetant △cgtf6 was used as the receptor strain.CRISPR-Cas9 gene knockout system was used to knock out Cgura3 gene encoding the orotidine 5’-phosphate decarboxylase and the ORF of Cgura3 was simultaneously replaced by yeGFP labeled CgTF6 gene.The promoter Ptf6 and Cgura3 promoter was used to  initiate the transcription of yeGFP-CgTF6 respectively.The promoter Ptf6 –yeGFP- CgTF6 fragment, the Cgura3 promoter–yeGFP- CgTF6 fragment as well as Cas9 expression vector were obtained by seamless ligation, TA cloning and overlapping PCR.QRT-PCR was employed to analyze the effect of epigenetic modification drugs on the transcription of Cgtf6 in the wild-type strain.The results showed that the histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) significantly inhibited the transcription of Cgtf6，and the mRNA level of Cgtf6 was reduced to The results showed that the histone deacetylase inhibitor SAHA (suberoylanilide hydroxamic acid) inhibited the transcription of CgTF6, and the mRNA level was down regulated to 30% of that in the wild-type strain. The results of this paper provide some vectors and experimental data for the functional study of CgTF6.