Scherer S, Davis R W
Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951-5. doi: 10.1073/pnas.76.10.4951.
We have developed a method that may be of general application for the stable introduction of foreign sequences or deletions, constructed in vitro, into the chromosomes of Saccharomyces cerevisiae. No vector sequences are present in the final strains. Ability to transform cells with DNA, availability of a single selective marker, and integration of the transforming DNA by homologous recombination into the chromosomes are the requirements of the system. Any isolated gene can be deleted or altered and then be used to replace the wild-type chromosomal copy. An internal deletion mutant of the his3 gene and a transposition of a galactose-inducible region into chromosome XV have been generated by using the ura3 gene as the selective marker.
我们已经开发出一种方法，该方法可能具有普遍适用性，可将体外构建的外源序列或缺失片段稳定导入酿酒酵母的染色体中。最终菌株中不存在载体序列。用DNA转化细胞的能力、单一选择标记的可用性以及通过同源重组将转化DNA整合到染色体中是该系统的要求。任何分离的基因都可以被删除或改变，然后用于取代野生型染色体拷贝。通过使用ura3基因作为选择标记，已产生了his3基因的内部缺失突变体以及将半乳糖诱导区域转座到第十五号染色体中。
