玉米叶片酵母双杂交cDNA文库的构建及评价  

Construction and Appreciation of Yeast Two Hybrid cDNA Library of Maize Leaf

作　　者：张军杰[1] 崔红军[2] 黄玉碧[2] 

机构地区：[1]四川农业大学理学院,四川雅安625014 [2]四川农业大学玉米研究所,四川雅安625014

出　　处：《西北农业学报》2008年第5期162-165,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家863计划项目(SQ2007AA10Z135024);四川省教育厅青年基金项目(2006B010)资助

摘　　要：以玉米骨干自交系478为试验材料,从各生长时期的叶片中提取总RNA后,再利用磁珠法从中分离纯化出mRNA,然后以mRNA为模板,反转录合成cDNA第一链,再在DNA聚合酶作用下,通过长距离PCR,扩增双链cDNA(ds cDNA)。利用SMART技术,通过同源重组的方法,在酵母菌株AH109中构建了玉米叶片全长cDNA文库。经检测,文库的转化率为1.54×107转化子/3μg pGADT7-Rec,文库滴度为1.03×106pfu/mL,重组率为95%。The maize inbred line 478 was used experiment material,and the total RNA was isolated from each periods of growth maize leaf and mRNA was separated and purified using the SA-PMPs method.Afterwords,mRNA was used as cyclostyle and synthesized the primacy cDNA.Under DNA polymerase function,DNA was expand double strand DNA by using LD-PCR approach.Using SMART technique and homologous reorganization method,the yeast two hybrid cDNA library was constructed in yeast AH109,After examination,the library conversion rate was 1.54×107 transfers beggar /3 μg pGADT7-Rec,the capacity of the library was 1.03×106 pfu/mL,and the reorganization rate was 95%.

关 键 词：玉米叶片 酵母双杂交文库 构建 

分 类 号：S513[农业科学—作物学] Q943.2[生物学—植物学]