Macreadie I G, Jagadish M N, Azad A A, Vaughan P R
CSIRO, Division of Biotechnology, Parkville, Victoria, Australia.
Plasmid. 1989 Mar;21(2):147-50. doi: 10.1016/0147-619x(89)90059-0.
A series of yeast expression vectors and cassettes utilizing the CUP1 gene of Saccharomyces cerevisiae have been constructed. The cassettes contain multiple cloning sites for gene fusions and were created by inserting a 27-bp polylinker at the +14 position of the CUP1 gene. The cassettes are portable as restriction fragments and enable copper-regulated expression of foreign proteins in S. cerevisiae. In copper sensitive yeast, multiple copies of the CUP1 cassettes confer copper resistance due to the production of the copper metallothionein. Genes cloned into the CUP1 cassettes, however, usually prevent translation of the metallothionein leading to a loss of resistance. This could be useful for one-step cloning into yeast.
已经构建了一系列利用酿酒酵母CUP1基因的酵母表达载体和盒式结构。这些盒式结构包含用于基因融合的多个克隆位点，是通过在CUP1基因的+14位置插入一个27 bp的多克隆位点而创建的。这些盒式结构可作为限制性片段进行转移，并能够在酿酒酵母中实现外源蛋白的铜调节表达。在对铜敏感的酵母中，由于铜金属硫蛋白的产生，多个拷贝的CUP1盒式结构赋予铜抗性。然而，克隆到CUP1盒式结构中的基因通常会阻止金属硫蛋白的翻译，导致抗性丧失。这对于一步克隆到酵母中可能是有用的。
