Unkles S E, Campbell E I, Carrez D, Grieve C, Contreras R, Fiers W, Van den Hondel C A, Kinghorn J R
Plant Molecular Genetics Unit, University of St. Andrews, Fife, U.K.
Gene. 1989 May 15;78(1):157-66. doi: 10.1016/0378-1119(89)90323-5.
A homologous transformation for Aspergillus niger was developed based on the nitrate reductase structural gene niaD. This system offered certain advantages over existing A. niger systems, such as the ease of recipient mutant isolation, absence of abortive transformants, convenient enzyme assay, ease of transformant stability testing, and complete absence of background growth. Transformation frequencies of up to 100 transformants per microgram DNA were obtained with the vector pSTA10 which carries the niaD gene of A. niger. Southern blotting analysis indicated that vector DNA had integrated into the genome of A. niger. Mitotic stability studies demonstrated that while some transformants were as stable as the wild-type (wt), others were markedly less so. No correlation was seen between plasmid integration, mitotic stability and nitrate reductase activity, which was markedly different from wt in only three of the transformants examined.
基于硝酸还原酶结构基因niaD开发了一种黑曲霉的同源转化方法。该系统相对于现有的黑曲霉系统具有某些优势，例如易于分离受体突变体、不存在流产转化体、酶分析方便、转化体稳定性测试容易以及完全没有背景生长。使用携带黑曲霉niaD基因的载体pSTA10，获得了每微克DNA高达100个转化体的转化频率。Southern印迹分析表明载体DNA已整合到黑曲霉的基因组中。有丝分裂稳定性研究表明，虽然一些转化体与野生型一样稳定，但其他转化体的稳定性明显较差。在所检测的转化体中，只有三个转化体的质粒整合、有丝分裂稳定性与硝酸还原酶活性之间没有相关性，这与野生型明显不同。
