大白菜开花相关基因FLC1的BAC克隆筛选及分析

Screening and Analysis of BAC Clones Containing Flowering Time Gene FLC1 in Brassica campestris L. ssp. pekinensis Olsson

采用改良的混合池构建方法构建大白菜BAC文库一级混合池和二级混合池,利用开花相关基因FLC1特异引物对其进行PCR筛选。通过三步PCR扩增、114个PCR反应,筛选了19200个克隆,获得了2个FLC1单克隆。克隆测序结果表明,其扩增产物的序列与大白菜FLC1的相似性达到99%,证实此克隆为含FLC1基因的BAC克隆。

The primary and secondary pools of a bacterial artificial chromosome（BAC）library in Chinese cabbage were constructed using improved BAC pooling strategy.The specific primers of flowering time gene FLC1 were to screen those BAC clones by PCR amplification.In total two positive FLC1 clones were obtained from 19 200 BACs through three steps of PCR screening with 114 reactions.Sequence analysis of FLC1 clones showed that these fragments has 99% of homology with the FLC1 in Chinese cabbage,indicating that the flowering time gene FLC1 is contained in the BAC clones.