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[关于因多瘤病毒DNA片段而在酿酒酵母中复制的杂种质粒稳定性的研究]

[Study of the stability of hybrid plasmids replicating in Saccharomyces cerevisiae due to DNA fragments from polyoma virus].

作者信息

Oganesian N A, Chepurnoĭ A I, Vel'kov V V, Kopylova-Sviridova T N, Fodor I I

出版信息

Mol Biol (Mosk). 1984 Jan-Feb;18(1):21-9.

PMID:6323973
Abstract

Hybrid plasmid pSP97 carrying the entire genome of polyoma virus (PY), inserted into bacterial vector psV3, transforms yeast cells with the frequency 1 x 10(-2). Plasmid pSP97 is capable of autonomous replication in S. cerevisiae, while its structure remains unaltered, the stability of hybrid plasmid in transformants is 44%--100%. Plasmid pSP155 consisting of Ori-containing DNA segment from polyoma, pBR322 and yeast gene arg4, transforms yeast cells with the frequency 5 x 10(-3), the stability of plasmid in transformants is 23%--29%. Two types of plasmids were isolated from transformants: one was identical to SP155, while the another differed structurally and phenotypically from SP155. Plasmids pSP113 and pSP114, in addition to pBR322 and yeast gene arg4, contain a viral DNA segment that encodes genes from small and middle T-antigens. These plasmids transform yeast cells with low frequency (2 x 10(-4), 3 x 10(-5)), the stability of plasmids in yeast transformants is 100%. However, hybrid plasmids identical to pSP113 were isolated from transformants. Structural rearrangements have been observed in pSP114, which carries the arg4 gene in reversed orientation compared to pSP113.

摘要

携带多瘤病毒(PY)全基因组并插入细菌载体psV3的杂种质粒pSP97,以1×10(-2)的频率转化酵母细胞。质粒pSP97能够在酿酒酵母中自主复制,而其结构保持不变,杂种质粒在转化子中的稳定性为44%--100%。由来自多瘤病毒的含Ori DNA片段、pBR322和酵母基因arg4组成的质粒pSP155,以5×10(-3)的频率转化酵母细胞,质粒在转化子中的稳定性为23%--29%。从转化子中分离出两种类型的质粒:一种与SP155相同,而另一种在结构和表型上与SP155不同。质粒pSP113和pSP114,除了pBR322和酵母基因arg4外,还包含一个编码小T抗原和中T抗原基因的病毒DNA片段。这些质粒以低频率(2×10(-4),3×10(-5))转化酵母细胞,质粒在酵母转化子中的稳定性为100%。然而,从转化子中分离出了与pSP113相同的杂种质粒。在pSP114中观察到了结构重排,与pSP113相比,它携带的arg4基因方向相反。

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