Shervington A A, Sparey J A, Bostock C J
Agricultural and Food Research Council, Pirbright Laboratory, Woking, Surrey, U.K.
Biochem Mol Biol Int. 1993 Mar;29(4):673-85.
Plasmid, which combined the complete genome of BPV-I, yeast ARS, LEU yeast selectable marker gene, the NEO selectable marker gene and inverted (C4A2)n telomeric repeat gene sequences cloned originally from Tetrahymena thermophila, was constructed. It was introduced in either circular or linear form to Saccharomyces cerevisiae or Schizosaccharomyces pombe. Although both yeasts could replicate the plasmid extrachromosomally, irrespective of whether it was introduced as a circular or linear structure, the yeasts did differ in their ability to resolve a circular plasmid carrying the telomeric sequences into linear forms. S. cerevisiae was found to resolve the circular form of pCA/LEU/ARS to the linear structure, whereas circular pCA/LEU/ARS remained circular in S. pombe. On the other hand, pCA/LEU/ARS which had been previously linearised at the telomeric sequence was maintained as a linear structure in S. pombe and S. cerevisiae.
构建了一种质粒,其组合了BPV-I的完整基因组、酵母自主复制序列(ARS)、亮氨酸酵母选择标记基因、新霉素选择标记基因以及最初从嗜热四膜虫克隆的反向(C4A2)n端粒重复基因序列。该质粒以环状或线性形式导入酿酒酵母或粟酒裂殖酵母。尽管两种酵母都能在染色体外复制该质粒,无论其是以环状还是线性结构导入,但酵母在将携带端粒序列的环状质粒转化为线性形式的能力上存在差异。发现酿酒酵母能将pCA/LEU/ARS的环状形式转化为线性结构,而在粟酒裂殖酵母中,环状pCA/LEU/ARS仍保持环状。另一方面,先前在端粒序列处线性化的pCA/LEU/ARS在粟酒裂殖酵母和酿酒酵母中均维持线性结构。