携带酵母trp1区域DNA的质粒pBR313在酿酒酵母中的复制。

Replication in Saccharomyces cerevisiae of plasmid pBR313 carrying DNA from the yeast trpl region.

作者信息

Kingsman A J, Clarke L, Mortimer R K, Carbon J

出版信息

Gene. 1979 Oct;7(2):141-52. doi: 10.1016/0378-1119(79)90029-5.

Abstract

Plasmid pBR313 carrying a 1.4 kb EcoRI fragment from the yeast TRP1 region (designated pLC544) is capable of transforming yeast trp1 mutants to Trp+ at high frequency (10(3)--10(4) transformants/micrograms DNA). Transformation can be achieved either by using purified plasmid DNA or by fusion of yeast spheroplasts with partially lysed Escherichia coli [pLC544] protoplast preparations. The Trp+ yeast transformants are highly unstable, segregating Trp- cells at frequencies of 0.18 per cell per generation (haploids) and 0.056 per cell per generation (diploids) in media containing tryptophan. Plasmid pLC544 replicates autonomously in the nucleus of yeast cells and segregation of Trp-cells is associated with the complete loss of plasmid sequences. In genetic crosses, pLC544 is randomly assorted during meiosis and is carried unchanged through the mating process into haploid recombinants.

摘要

携带来自酵母TRP1区域的1.4 kb EcoRI片段的质粒pBR313（命名为pLC544）能够以高频率（10³ - 10⁴转化子/微克DNA）将酵母trp1突变体转化为Trp⁺。转化可以通过使用纯化的质粒DNA来实现，也可以通过酵母原生质球与部分裂解的大肠杆菌[pLC544]原生质体制备物融合来实现。Trp⁺酵母转化体高度不稳定，在含有色氨酸的培养基中，单倍体细胞每代以0.18的频率、二倍体细胞每代以0.056的频率分离出Trp⁻细胞。质粒pLC544在酵母细胞核中自主复制，Trp⁻细胞的分离与质粒序列的完全丢失相关。在遗传杂交中，pLC544在减数分裂过程中随机分配，并在交配过程中保持不变地传递到单倍体重组体中。