激活标记文库构建及突变体表型分析

Cao Dongmei, Fan Xi-Ying, Wang Yunshan, Kang Lifang
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引用次数: 1

摘要

激活标记在植物基因组学研究中起着重要的作用。本文通过介导转化拟南芥(Aribidopsis thaliana)突变体bzr1-D,构建了包含5万株抗basta细胞系的激活标记文库。其中47个转化品系表现出明显的表型,包括开花时间晚、生长习性矮、叶形改变、叶柄变长、缺毛、不育、茎叶间无扭结等。利用热不对称交错PCR和巢式PCR获得部分T-DNA侧翼序列。结果显示,突变体含有1 - 3个T-DNA插入。这些插入物分布在基因组的第1、4和5条染色体上。
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Activation tagging library construction and mutant phenotype analysis
Activation tagging plays an important role in plant genomics studies. In this paper, an activation tagging library containing 50 000 Basta-resistant lines was constructed by mediated transformation of the mutant bzr1-D of Aribidopsis thaliana . Among these transformants, 47 lines showed obvious phenotypes, including late flowering time, dwarf growth habit, changed leaf shape, longer leaf petiole, leaves lacking trichomes, sterility and no kink between stem/leaf. Some T-DNA flanking sequences were obtained by TAIL (thermal asymmetric interlaced) and nested PCR. Results showed that the mutants contained one to three T-DNA insertions. The insertions were distributed in the first, fourth and fifth chromosomes of the genome.
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