塔宾曲霉的鉴定及基因编辑方法对其同源重组效率的影响

打开文本图片集

中图分类号：Q78 文献标志码：A 文章编号：1008-0864（2025）06-0083-10

Identification of Aspergillus tubingensis and the Effect ofGene Editing Methods on Its Homologous Recombination Efficiency

LIANG Licun1，YU Huimin1，GUAN Xiaochen²，HUANG Huoqing¹，YAO Bin1，YANG Haomengl* （1.InstituteofAnimalSciences，ChineseAcademyofAgriculturalSienes，BeijingO193，China；2.GannanCountyQualityn Technical Supervision Inspection and Testing Center，Heilongjiang Qiqihar 162199，China）

Abstract：Aspergilustubingensisisafilamentousfungus withbroadapplicationprospectsinfeed，food，biological control，environmentalrestorationand soon.Combining morphologicalobservation withmoleculardetection，Ehrlich reagentstaining，a strain of Aspergillus tubingensis was succefullyidentifiedand was named asF316.Inorder to explore asuitable gene editing method for F316 and facilitate genetic enginering modification，CRISPR/Cas9 was applied to insert hygromycin resistance genes into the spore pigment synthesis related gene funA.Through spore color statisticsandresistance genePCRverification，theeffctsof different delivery methodsof Cas9andgRNAon homologous recombination eficiency were compared.Theresults showed that，for F316 strain，the homologous recombination eficiencyof expression cassettemethod，self-replicating plasmid method and ribonucleoprotein complex（RNP）method were 70.14% ，66.67%and 25.70% ，respectively.Inaddition，the length of homologous arms in donor DNA also affcted the homologous recombination eficiency.Whenthe length of homologous arms was 2000bp ，the recombination efficiency was the best.

Key Words：Aspergillus tubingensis；strain identification；gene editing；homologous recombination

塔宾曲霉（Aspergillustubingensis）是子囊菌纲、曲霉属真菌，在食品发酵、生物防治、环境污染治理等方面具有广泛的应用前景。（剩余14056字）