Van Lijsebettens M, Wang X, Cnops G, Boerjan W, Desnos T, Höfte H, Van Montagu M
Laboratorium voor Genetica, VIB Universiteit Gent, Belgium.
Mol Gen Genet. 1996 Jun 12;251(3):365-72. doi: 10.1007/BF02172528.
The map positions of a set of eight T-DNA insertions in the Arabidopsis genome have been determined by using closely linked visible markers. The insertions are dispersed over four of the five chromosomes. Each T-DNA insert contains one or more of the chimeric marker genes neomycin phosphotransferase (neo), hygromycin phosphotransferase (hpt), phosphinothricin acetyltransferase (bar), beta-glucuronidase (gusA) and indole-3-acetamide hydrolase (iaaH). The neo, hpt and bar marker genes are dominant in a selective germination assay or when used as DNA markers in a polymerase chain reaction. These dominant markers will allow recombinants to be discerned in a germinating F2 population, one generation earlier than with a conventional recessive marker. The transgenic marker lines will speed up and simplify the isolation of recombinants in small genetic intervals, a rate-limiting step in positional cloning strategies. The transgenic lines containing the hpt marker will also be of interest for the isolation of deletion mutants at the T-DNA integration sites.
利用紧密连锁的可见标记,已确定了拟南芥基因组中一组8个T-DNA插入位点的图谱位置。这些插入分布在五条染色体中的四条上。每个T-DNA插入片段包含一个或多个嵌合标记基因,如新霉素磷酸转移酶(neo)、潮霉素磷酸转移酶(hpt)、膦丝菌素乙酰转移酶(bar)、β-葡萄糖醛酸酶(gusA)和吲哚-3-乙酰胺水解酶(iaaH)。在选择性萌发试验中,或用作聚合酶链反应中的DNA标记时,neo、hpt和bar标记基因具有显性。这些显性标记将使重组体在F2萌发群体中得以辨别,比使用传统隐性标记早一代。转基因标记系将加快并简化小遗传区间内重组体的分离,这是定位克隆策略中的一个限速步骤。含有hpt标记的转基因系对于分离T-DNA整合位点处的缺失突变体也将具有重要意义。