Transformation of the nematode-trapping fungus Arthrobotrys oligospora
Transformation of the nematode-trapping fungus Arthrobotrys oligospora
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DOI：
10.1111/j.1574-6968.1999.tb13491.x
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发表时间：
1999-04-01
影响因子：
2.1
通讯作者：
Oliver, RP
中科院分区：
文献类型：
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作者：
Tunlid, A;Åhman, J;Oliver, RP
利用来自大肠杆菌的潮霉素B磷酸转移酶基因，在多种异源真菌启动子的控制下，将捕食线虫的真菌少孢节丛孢菌转化为潮霉素抗性菌株。用聚乙二醇/CaCl2处理，将质粒DNA导入真菌原生质体。转化频率在1-6个转化子/亩DNA之间。从转化子中分析的13个整合事件中有7个是单拷贝整合，而其余的是多个和更复杂的整合。在转化过程中加入限制性内切酶提高了单拷贝整合的频率。在尼杜拉曲霉启动子的控制下，利用编码β-葡萄糖苷酸酶报告基因的大肠杆菌uidA基因进行共转化，转化频率高达63%。(C)1999年欧洲微生物学会联合会。爱思唯尔科学公司出版。版权所有。
The nematode-trapping fungus Arthrobotrys oligospora was transformed to hygromycin resistance using the hygromycin-B phosphotransferase gene from Escherichia coli under the control of various heterologous fungal promoters. Plasmid DNA was introduced into fungal protoplasts by polyethylene glycol/CaCl2 treatment. Transformation frequencies varied between 1-6 transformants per mu g DNA. Seven out of 13 integration events analyzed from transformants were single copy integrations, whereas the remaining were multiple and more complex integrations. The addition of restriction enzymes during transformations increased the frequency of single copy integrations. Co-transformation, using the E. coli uidA gene encoding the beta-glucuronidase reporter gene under the control of an Aspergillus nidulans promoter, occurred at frequencies of up to 63%. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.