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大型拟南芥海甘蓝 BIBAC 插入文库的基因组学研究和耐逆基因的鉴定。

A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes.

机构信息

State Key Laboratory for Biocontrol, Sun Yat-sen (Zhongshan) University, 510275, Guangzhou, China.

出版信息

Plant Mol Biol. 2010 Jan;72(1-2):91-9. doi: 10.1007/s11103-009-9553-3. Epub 2009 Sep 29.

Abstract

Salt cress (Thellungiella halophila), a salt-tolerant relative of Arabidopsis, has turned to be an important model plant for studying abiotic stress tolerance. One binary bacterial artificial chromosome (BIBAC) library was constructed which represents the first plant-transformation-competent large-insert DNA library generated for Thellungiella halophila. The BIBAC library was constructed in BamHI site of binary vector pBIBAC2 by ligation of partial digested nuclear DNA of Thellungiella halophila. This library consists of 23,040 clones with an average insert size of 75 kb, and covers 4x Thellungiella halophila haploid genomes. BIBAC clones which contain inserts over 50 kb were selected and transformed into Arabidopsis for salt tolerant plant screening. One transgenic line was found to be more salt tolerant than wild type plants from the screen of 200 lines. It was demonstrated that the library contains candidates of stress tolerance genes and the approach is suitable for the transformation of stress susceptible plants for genetic improvement.

摘要

盐地碱蓬(Thellungiella halophila),一种耐盐的拟南芥近缘植物,已成为研究非生物胁迫耐受性的重要模式植物。构建了一个双元细菌人工染色体(BIBAC)文库,这是第一个为盐地碱蓬生成的具有转化能力的大型插入 DNA 文库。BIBAC 文库通过将盐地碱蓬的部分消化核 DNA 连接到二元载体 pBIBAC2 的 BamHI 位点而构建。该文库包含 23,040 个克隆,平均插入大小为 75 kb,覆盖了盐地碱蓬单倍体基因组的 4 倍。选择包含超过 50 kb 插入片段的 BIBAC 克隆,并将其转化为拟南芥进行耐盐植物筛选。从 200 个品系的筛选中发现,一个转基因株系比野生型植物更耐盐。结果表明,该文库包含耐胁迫基因的候选基因,并且该方法适用于遗传改良中对胁迫敏感植物的转化。

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