家蚕耐氟基因RAPD分子标记的筛选及其克隆

Screening and Cloning of RAPD Marker of Fluoride Tolerance Gene in Silkworm(Bombyx mori)

为获得家蚕耐氟基因的分子标记 ,以家蚕耐氟品种T6和高敏感品种 733新为材料 ,并构建了其近等基因系 ,采用 30 0个随机引物进行RAPD扩增 ,获得了与家蚕耐氟性有关的一个分子标记 ,此标记由引物S2 5 0扩增得到 ,并在F2代分离个体中得到验证 ,证明了此分子标记的可靠性 ,进而将此标记克隆进T载体pMD - 18中 ,完成了测序 。

In order to find out the molecular marker that linked to the fluoride tolerance gene of the silkworm,we used silkworm strain T6 with tolerant to fluoride and silkworm strain 733xin with highly sensitive to fluoride to construct the near-isogenic lines.300 random primers were used in RAPD amplification to DNAs of these lines.A molecular marker amplified by S250 that linked to the fluoride tolerance gene was obtained.The molecular marker was examined and verified in F2 separated individuals.Subsequently,the molecular marker was cloned into T vector pMD-18 for sequencing.Comparing with sequences available in the GenBank showed that this molecular marker was novel.