Powell W A, Kistler H C
Plant Pathology Department, University of Florida, Gainesville 32611.
J Bacteriol. 1990 Jun;172(6):3163-71. doi: 10.1128/jb.172.6.3163-3171.1990.
Particular combinations of fungal strains and transformation vectors allow for fungal rearrangement of normally integrative plasmids, resulting in the creation of linear self-replicating plasmids in Fusarium oxysporum. The rearrangement results in the addition of fungal DNA, including telomere consensus sequences, to plasmid termini. The mechanism by which this rearrangement occurs is unclear, but it has similarities to extrachromosomal gene amplification. A DNA fragment which allows for linear autonomous replication upon reintroduction to the fungus was subcloned and sequenced. This DNA sequence contains the repeated telomeric sequence TTAGGG flanked by a region of twofold symmetry consisting primarily of pUC12 DNA. Isolation and identification of this sequence is the first step toward development of vectors that function as artificial chromosomes in filamentous fungi. This sequence was shown to promote autonomous replication and enhance transformation in several strains of F. oxysporum, Nectria haematococca, and Cryphonectria parasitica.
特定的真菌菌株和转化载体组合可使正常整合型质粒在真菌中发生重排,从而在尖孢镰刀菌中产生线性自我复制质粒。这种重排导致真菌DNA(包括端粒共有序列)添加到质粒末端。这种重排发生的机制尚不清楚,但它与染色体外基因扩增有相似之处。一个在重新导入真菌后能实现线性自主复制的DNA片段被亚克隆并测序。该DNA序列包含重复的端粒序列TTAGGG,两侧是主要由pUC12 DNA组成的双重对称区域。分离和鉴定该序列是开发在丝状真菌中起人工染色体作用的载体的第一步。该序列在几种尖孢镰刀菌、血红红球菌和寄生隐孢壳菌菌株中均显示出促进自主复制和增强转化的作用。