甘蓝基因组细菌人工染色体文库的构建

Construction of a Bacterial Artificial Chromosome Library from Brassica oleracea

细菌人工染色体(BAC)文库是克隆甘蓝抗病优质重要基因的基础,以抗枯萎病、富含硫代葡萄糖苷的结球甘蓝自交系R4P1为材料,美国Epicentre公司的CopyControlTMpCC1BACTM为载体,构建了甘蓝细菌人工染色体文库。该文库有73 344个克隆,插入片段平均大小约97 kb,空载率<3%,覆盖甘蓝基因组约10.9倍,筛选到甘蓝任一基因的概率为99.99%,这些结果表明,构建的文库质量较好,可用于后续分析。构建的甘蓝BAC文库不仅可用于枯萎病基因的克隆,而且为克隆其他重要的功能基因及基因组学等的研究奠定了基础。

Bacterial artificial chromosome library is the foundation of important and high quality disease-resistant gene cloning from Brassica oleracea. A bacterial artificial chromosome library was constructed from R4P1,a Brassica oleracea varieties highly resisting to fusarium wilt and being in rich of glucosinolates,with the vector CopyControlTMpCC1 BACTM. The library consists of 73 344 clones with an average insert size of about 97 kb,and in which the empty clones are under 3%. Thus,the library provides 10. 9 haploid genome equivalents,the probability of screening to any gene of B. oleracea is 99. 99%,these results indicate that this is a high quality BAC library suitable for the following analysis. The BAC library of B. oleracea could not only be used for the clone of resistant genes to fusarium wilt,but also lay foundations for isolating other interesting genes and research the genomics of B. oleracea,et al.