玉米胚芽鞘负向重力生长时期的酵母双杂交文库构建及评价

Construction and Appreciation of Yeast Two Hybrid Library of Maize Coleoptile during Negtive-gravitropic Growth

[目的]构建玉米胚芽鞘负向重力生长时期的酵母双杂交文库,并对其进行评价。[方法]以玉米骨干自交系B73为材料,利用Trizol试剂提取玉米胚芽鞘在负向生长时期所表达的总RNA,再利用SMART技术构建酵母双杂交文库,并转化进酵母Y187中。[结果]试验得到的原始文库库容量高达1×106个单克隆;文库中插入的片段集中在0.5～1.5 kb,平均插入长度为800 bp。[结论]试验构建得到了高质量的米胚芽鞘酵母双杂交文库,为研究胚芽鞘的负向重力反应机制奠定了基础。

[Objective ] The aim was to construct and appraise the yeast two hybrid library of maize coleoptile during its negative-gravitropie growth. [ Method] Using maize inbred line B73 as materials, the total RNA which expressed during negative-gravitropic growth period of maize coleoptile was extracted by Trizol, and the yeast two hybrid library was constructed by SMART technology and transformed into yeast Y187.[ Result] The results showed that the library contained 1× 10^6 monoelones and the predominant size of inserted fragments in library was 0.5 - 1.5 kh, average size was 800 bp. [ Conelusion] High-quality yeast two hybrid library of maize eoleoptile was constructed and it prepared the ground for the study of negative-gravitropie reaction mechanism of eoleopfile.