Institut für Genetik, Universität zu Köln, Weyertal 121, D-5000 Köln 41, FRG.
EMBO J. 1982;1(11):1455-60. doi: 10.1002/j.1460-2075.1982.tb01337.x.
In an attempt to isolate the transposable genetic element Ds from Zea mays L., we cloned DNA fragments hybridizing to a cDNA clone derived from the sucrose synthase gene in a lambda vector (lambda::Zm Sh). The fragments cloned from wild-type and from the Ds-induced mutant sh-m5933 (lambda::Zm sh-m5933) share a segment 6 kb long while a contiguous segment of 15 kb of lambda::Zm sh-m5933 (mutant-derived DNA) does not hybridize to the DNA segment cloned from the wild-type. Restriction maps are given, and the junction point between the two DNA segments in the mutant clone was determined. Hybridization of DNA fragments, present in the wild-type DNA of lambda::Zm Sh, but not in the mutant clone, lambda::Zm sh-m5933, to genomic DNA of sh-m5933 showed that no part of this DNA is deleted. It cannot be said whether the DNA found in the mutant, but not in the wild-type clone, has been brought there by Ds insertion or by another Ds-dependent DNA rearrangement. The mutant-derived DNA was hybridized to genomic DNA of various maize lines digested by several restriction endonucleases. Approximately 40 bands were detected. The mutant-derived DNA contains two pairs of inverted repeats several hundred nucleotide pairs long, one of which is located at the junction to wild-type-derived DNA.
为了从玉米中分离转座遗传因子 Ds,我们将与蔗糖合酶基因 cDNA 克隆杂交的 DNA 片段克隆到λ载体(λ::Zm Sh)中。从野生型和 Ds 诱导的突变体 sh-m5933(λ::Zm sh-m5933)中克隆的片段共享 6kb 长的片段,而 15kb 长的连续片段λ::Zm sh-m5933(突变体衍生 DNA)与从野生型克隆的 DNA 片段不杂交。给出了限制图谱,并确定了突变体克隆中两个 DNA 片段之间的连接点。存在于λ::Zm Sh 的野生型 DNA 中但不存在于突变体克隆λ::Zm sh-m5933 中的 DNA 片段与 sh-m5933 的基因组 DNA 的杂交表明,没有部分 DNA 缺失。不能说突变体中发现的而不是在野生型克隆中发现的 DNA 是由 Ds 插入或其他 Ds 依赖的 DNA 重排带来的。突变体衍生的 DNA 与用几种限制内切酶消化的各种玉米品系的基因组 DNA 杂交。检测到大约 40 个带。突变体衍生的 DNA 包含两对长几百个核苷酸对的反向重复序列,其中一个位于与野生型衍生 DNA 的连接处。