Fincham J R
Microbiol Rev. 1989 Mar;53(1):148-70. doi: 10.1128/mr.53.1.148-170.1989.
Transformation with exogenous deoxyribonucleic acid (DNA) now appears to be possible with all fungal species, or at least all that can be grown in culture. This field of research is at present dominated by Saccharomyces cerevisiae and two filamentous members of the class Ascomycetes, Aspergillus nidulans and Neurospora crassa, with substantial contributions also from fission yeast (Schizosaccharomyces pombe) and another filamentous member of the class Ascomycetes, Podospora anserina. However, transformation has been demonstrated, and will no doubt be extensively used, in representatives of most of the main fungal classes, including Phycomycetes, Basidiomycetes (the order Agaricales and Ustilago species), and a number of the Fungi Imperfecti. The list includes a number of plant pathogens, and transformation is likely to become important in the analysis of the molecular basis of pathogenicity. Transformation may be maintained either by using an autonomously replicating plasmid as a vehicle for the transforming DNA or through integration of the DNA into the chromosomes. In S. cerevisiae and other yeasts, a variety of autonomously replicating plasmids have been used successfully, some of them designed for use as shuttle vectors for Escherichia coli as well as for yeast transformation. Suitable plasmids are not yet available for use in filamentous fungi, in which stable transformation is dependent on chromosomal integration. In Saccharomyces cerevisiae, integration of transforming DNA is virtually always by homology; in filamentous fungi, in contrast, it occurs just as frequently at nonhomologous (ectopic) chromosomal sites. The main importance of transformation in fungi at present is in connection with gene cloning and the analysis of gene function. The most advanced work is being done with S. cerevisiae, in which the virtual restriction of stable DNA integration to homologous chromosome loci enables gene disruption and gene replacement to be carried out with greater precision and efficiency than is possible in other species that show a high proportion of DNA integration events at nonhomologous (ectopic) sites. With a little more trouble, however, the methodology pioneered for S. cerevisiae can be applied to other fungi too. Transformation of fungi with DNA constructs designed for high gene expression and efficient secretion of gene products appears to have great commercial potential.
现在看来，用外源脱氧核糖核酸（DNA）对所有真菌物种进行转化都是可能的，或者至少对所有能够在培养条件下生长的真菌物种是可行的。目前，该研究领域主要由酿酒酵母以及子囊菌纲的两个丝状成员——构巢曲霉和粗糙脉孢菌主导，裂殖酵母（粟酒裂殖酵母）和子囊菌纲的另一个丝状成员——雪白粪壳菌也做出了重要贡献。然而，在大多数主要真菌类别的代表中，包括藻状菌纲、担子菌纲（伞菌目和黑粉菌属物种）以及一些半知菌类，都已经证明了可以进行转化，并且毫无疑问将会被广泛应用。这个名单中包括一些植物病原体，转化在致病性分子基础的分析中可能会变得很重要。转化可以通过使用自主复制质粒作为转化DNA的载体来维持，也可以通过将DNA整合到染色体中来实现。在酿酒酵母和其他酵母中，已经成功使用了多种自主复制质粒，其中一些被设计用作大肠杆菌的穿梭载体以及用于酵母转化。目前还没有适用于丝状真菌的合适质粒，在丝状真菌中，稳定转化依赖于染色体整合。在酿酒酵母中，转化DNA的整合几乎总是通过同源性进行；相比之下，在丝状真菌中，它在非同源（异位）染色体位点发生的频率同样很高。目前，转化在真菌中的主要重要性与基因克隆和基因功能分析有关。最先进的工作是在酿酒酵母中进行的，在酿酒酵母中，稳定DNA整合几乎局限于同源染色体位点，这使得基因破坏和基因替换能够比在其他在非同源（异位）位点有高比例DNA整合事件的物种中更精确、更高效地进行。然而，稍微多费点事，为酿酒酵母开创的方法也可以应用于其他真菌。用设计用于高基因表达和高效分泌基因产物的DNA构建体对真菌进行转化似乎具有巨大的商业潜力。
