Nyilasi Ildikó, Papp Tamás, Csernetics Arpád, Vágvölgyi Csaba
Department of Microbiology, Faculty of Sciences, University of Szeged, H-6701 Szeged, Hungary.
J Basic Microbiol. 2008 Feb;48(1):59-64. doi: 10.1002/jobm.200700221.
The Agrobacterium -mediated transformation was adapted to Backusella lamprospora, a zygomycete fungus closely related to Mucor. The transforming plasmid contained the hygromycin B resistance (hph) and the green fluorescent protein (gfp) genes under the control of the regulator regions of the Mucor circinelloides gpd1 gene. The presence of the hph and gfp genes in the transformants was detected by PCR. The introduced genes could also be amplified directly from the spores of the transformants. The transformation efficiency was investigated by fluorescence microscopy of the transformed spores. A gradual decrease in the hygromycin B resistance was observed during several cultivation cycles: the growth of the transformants on the selection medium became slower, and the detection of the introduced gene became more difficult.
根癌农杆菌介导的转化方法被应用于巴氏光孢霉，这是一种与毛霉密切相关的接合菌纲真菌。转化质粒包含潮霉素B抗性（hph）基因和绿色荧光蛋白（gfp）基因，它们受米根霉gpd1基因调控区的控制。通过聚合酶链反应（PCR）检测转化体中hph和gfp基因的存在情况。导入的基因也可以直接从转化体的孢子中扩增出来。通过对转化孢子进行荧光显微镜观察来研究转化效率。在几个培养周期中观察到潮霉素B抗性逐渐降低：转化体在选择培养基上的生长变得更慢，并且检测导入基因变得更加困难。
