塔賓曲霉的鑒定及基因編輯方法對(duì)其同源重組效率的影響

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中圖分類號(hào)：Q78 文獻(xiàn)標(biāo)志碼：A 文章編號(hào)：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

塔賓曲霉（Aspergillustubingensis）是子囊菌綱、曲霉屬真菌，在食品發(fā)酵、生物防治、環(huán)境污染治理等方面具有廣泛的應(yīng)用前景。（剩余14056字）