利用工程化Ds元件在玉米中进行基因标记

Gene tagging with engineered Ds elements in maize.

作者信息

Li Yubin, Segal Gregorio, Wang Qinghua, Dooner Hugo K

机构信息

Waksman Institute, Rutgers University, Piscataway, NJ, USA.

出版信息

Methods Mol Biol. 2013;1057:83-99. doi: 10.1007/978-1-62703-568-2_6.

DOI:10.1007/978-1-62703-568-2_6

PMID:23918422

Abstract

We describe here protocols for isolating genes in maize using Dissociation (Ds) transposons marked with a green fluorescent protein (GFP) transgene. The introduced marker enables the phenotypic scoring of the nonautonomous element and the anchoring of unique primers on the element to facilitate the isolation of the adjacent DNA by PCR. Transposons such as Ds transpose preferentially to sites closely linked to the Ds-launching platform. Based on this transposition behavior, a genetic resource is being created to mobilize a modified Ds element from different starting sites in the genome. Enough transgenic lines are being generated to cover most of the maize genome, allowing the targeted tagging of most genes from a Ds-launching platform located nearby.

摘要

我们在此描述了利用标记有绿色荧光蛋白（GFP）转基因的解离（Ds）转座子在玉米中分离基因的方法。引入的标记能够对非自主元件进行表型评分，并在该元件上锚定独特引物，以便通过聚合酶链反应（PCR）分离相邻的DNA。诸如Ds之类的转座子优先转座到与Ds起始平台紧密连锁的位点。基于这种转座行为，正在创建一种遗传资源，以从基因组中的不同起始位点动员修饰的Ds元件。正在产生足够数量的转基因系以覆盖玉米基因组的大部分，从而能够从附近的Ds起始平台对大多数基因进行靶向标记。