Hohmann S
Institut für Mikrobiologie,Technische Hochschule Darmstadt, Federal Republic of Germany.
Curr Genet. 1987;12(7):519-26. doi: 10.1007/BF00419561.
Integrative transformation of yeast with gapped DNA fragments results in single or multiple integration into the yeast genome via homologous recombination. A sequence of yeast DNA was found which favours multiple integration even when the strategy of gene replacement is used. This strategy by which the transformed DNA fragment replaces its chromosomal homologue rather than simply integrating into the genome usually occurs as a single exchange event. The described region is unique and lies near a telomere about 5 kb proximal to the SUC4 locus on chromosome XIII. DNA from this region was used as a vehicle for the integration of different SUC genes coding for invertase. Most of the sucrose fermenting transformants isolated carried between two and seven copies of the SUC genes. These transformants overproduced invertase even though there was no selective pressure for high invertase activity in these experiments. I conclude that this region is highly recombinogenic and favours multiple integration of DNA fragments. This region could be used for stable multiple integration of heterologous genes into the yeast genome for over-production of the respective gene product.
用缺口DNA片段对酵母进行整合转化会通过同源重组导致单拷贝或多拷贝整合到酵母基因组中。发现了一段酵母DNA序列，即使采用基因置换策略，该序列也有利于多拷贝整合。转化的DNA片段取代其染色体同源物而非简单地整合到基因组中的这种策略通常作为单交换事件发生。所描述的区域是独特的，位于第十三号染色体上SUC4基因座近端约5 kb处的一个端粒附近。来自该区域的DNA被用作整合不同编码转化酶的SUC基因的载体。分离得到的大多数蔗糖发酵转化子携带两到七个拷贝的SUC基因。尽管在这些实验中对高转化酶活性没有选择压力，但这些转化子仍过量产生转化酶。我得出结论，该区域具有高度的重组活性，有利于DNA片段的多拷贝整合。该区域可用于将异源基因稳定地多拷贝整合到酵母基因组中，以过量生产相应的基因产物。
