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用重组黏粒文库获得的粗糙脉孢菌inl+转化体的鉴定

Characterization of inl+ transformants of Neurospora crassa obtained with a recombinant cosmid-pool.

作者信息

Fehér Z, Schablik M, Kiss A, Zsindely A, Szabó G

机构信息

Department of Biology, University Medical School, Debrecen, Hungary.

出版信息

Curr Genet. 1986;11(2):131-7. doi: 10.1007/BF00378205.

Abstract

We constructed a Neurospora crassa gene library in a cosmid vector and used the cosmid-pool DNA to transform an inl, rg Neurospora crassa strain to inositol prototrophy. The inl+ colonies obtained in this experiment proved to be integrative type transformants. Genetic analysis revealed that the integration event occurred at or near the inl locus. In one of the transformants the inl+ trait exhibited mitotic and meiotic instability. In hybridization experiments free plasmids were detected in the F1 progeny of the transformants. We were able to recover eleven different plasmids from the F1 progeny of the transformants. None of these plasmids proved to carry a functional copy of the inl+ gene as judged by its transforming ability. Possible explanations for the observed phenomena are discussed.

摘要

我们构建了一个以黏粒载体为基础的粗糙脉孢菌基因文库,并使用黏粒池DNA将粗糙脉孢菌的inl、rg菌株转化为肌醇原养型。在该实验中获得的inl+菌落被证明是整合型转化体。遗传分析表明,整合事件发生在inl位点或其附近。在其中一个转化体中,inl+性状表现出有丝分裂和减数分裂不稳定性。在杂交实验中,在转化体的F1后代中检测到了游离质粒。我们能够从转化体的F1后代中回收11种不同的质粒。根据其转化能力判断,这些质粒均未携带inl+基因的功能拷贝。文中讨论了对观察到的现象的可能解释。

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