Fotheringham S, Holloman W K
Interdivisional Program in Molecular Biology, Graduate School of Medical Sciences, Cornell University Medical College, New York, New York 10021.
Mol Cell Biol. 1989 Sep;9(9):4052-5. doi: 10.1128/mcb.9.9.4052-4055.1989.
We have demonstrated that genes from Ustilago maydis can be cloned by direct complementation of mutants through the use of genomic libraries made in a high-frequency transformation vector. We isolated a gene involved in amino acid biosynthesis as an illustrative example and showed that integrative and one-step disruption methods can be used to create null mutations in the chromosomal copy of the gene by homologous recombination. The results of this investigation make it clear that one-step gene disruption will be of general utility in investigations of U. maydis, since simple, precise replacement of the sequence under study was readily achieved.
我们已经证明，通过使用高频转化载体构建的基因组文库，可以通过直接互补突变体来克隆来自玉米黑粉菌的基因。作为一个示例，我们分离出了一个参与氨基酸生物合成的基因，并表明整合和一步破坏方法可用于通过同源重组在该基因的染色体拷贝中产生无效突变。这项研究的结果清楚地表明，一步基因破坏在玉米黑粉菌的研究中将具有普遍用途，因为很容易实现对所研究序列的简单、精确替换。
