甘蓝型油菜基因组文库的构建及与硼高效基因相连锁克隆的筛选

以甘蓝型油菜宁RS-1为材料,构建了含有82944个克隆的甘蓝型油菜的BAC基因组文库.从文库中随机挑取克隆进行DNA长度检测,BAC克隆平均插入片段大小为80 kb,覆盖甘蓝型油菜基因组的5.1倍.随机挑取108克隆进行继代培养100代,分离质粒酶切检测表明不存在插入片段丢失现象,表明该文库的克隆在大肠杆菌中稳定存在;以与硼高效基因相连

Construction of a Brassica napus Bacterial Artificial Chromosome Library and Identification of Clones Linked to Boron Efficiency Gene

A bacterial artificial chromosome (BAC) library was constructed for a Brassica napus cultivar: Ning RS 1. The library contains 82944 clones with 80kb DNA insert size in average representing equivalents of 5 1 Brassica napus haploid genome. The BAC DNA can maintain stably in E. coli after about 100 generations of culture. pa28, an Arabidopsis EST clone which linked to a boron efficiency gene BE1 in B.napus , was used as probe to screen the library: consequently, 13 positive clones were identified. These positive clones were divided into four groups. Three of 13 clones were identified to link to the BE1 locus by Southern analysis.