Garza D, Medhora M, Koga A, Hartl D L
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110-1095.
Genetics. 1991 Jun;128(2):303-10. doi: 10.1093/genetics/128.2.303.
A chimeric white gene (wpch) and other constructs containing the transposable element mariner from Drosophila mauritiana were introduced into the germline of Drosophila melanogaster using transformation mediated by the P element. In the absence of other mariner elements, the wpch allele is genetically stable in both germ cells and somatic cells, indicating that the peach element (i.e., the particular copy of mariner inserted in the wpch allele) is inactive. However, in the presence of the active element Mos1, the wpch allele reverts, owing to excision of the peach element, yielding eye-color mosaics and a high rate of germline reversion. In strains containing Mos1 virtually every fly is an eye-color mosaic, and the rate of wpch germline reversion ranges from 10 to 25%, depending on temperature. The overall rates of mariner excision and transposition are approximately sixfold greater than the rates in comparable strains of Drosophila simulans. The activity of the Mos1 element is markedly affected by position effects at the site of Mos1 insertion. In low level mosiac lines, dosage effects of Mos1 are apparent in the heavier level of eye-color mosaicism in Mos1 homozygotes than in heterozygotes. However, saturation occurs in high level mosaic lines, and then dosage effects are not observed. A pBluescribe M13+ plasmid containing Mos1 was injected into the pole plasm of D. melanogaster embryos, and the Mos1 element spontaneously integrated into the germline at high efficiency. These transformed strains of D. melanogaster presently contain numerous copies of mariner and may be useful in transposon tagging and other applications.
利用P因子介导的转化方法,将一个嵌合白色基因(wpch)以及其他含有来自毛里求斯果蝇的转座元件mariner的构建体导入黑腹果蝇的种系中。在没有其他mariner元件的情况下,wpch等位基因在生殖细胞和体细胞中都是遗传稳定的,这表明桃元件(即插入wpch等位基因中的mariner的特定拷贝)是无活性的。然而,在存在活性元件Mos1的情况下,wpch等位基因会发生回复突变,这是由于桃元件的切除,产生眼色嵌合体和高频率的种系回复突变。在含有Mos1的品系中,几乎每只果蝇都是眼色嵌合体,并且wpch种系回复突变的频率在10%到25%之间,这取决于温度。mariner切除和转座的总体频率大约是拟暗果蝇可比品系中频率的六倍。Mos1元件的活性受到Mos1插入位点位置效应的显著影响。在低水平嵌合品系中,Mos1的剂量效应在Mos1纯合子比杂合子中更重的眼色嵌合水平上明显可见。然而,在高水平嵌合品系中会出现饱和现象,然后就观察不到剂量效应了。将一个含有Mos1的pBluescribe M13 + 质粒注射到黑腹果蝇胚胎的极质中,Mos1元件会高效地自发整合到种系中。这些转化的黑腹果蝇品系目前含有大量mariner拷贝,可能在转座子标签及其他应用中有用。