|
|||||
|
|
| 花生蛴螬中肠组织cDNA文库的构建 | |
| 引用本文： | 刘小民,张霞,李杰,郭巍.花生蛴螬中肠组织cDNA文库的构建[J].花生学报,2010,39(1):15-19. |
| 作者姓名： | 刘小民 张霞 李杰 郭巍 |
| 作者单位： | 1. 河北农业大学植物保护学院,河北,保定,071001 2. 河北省科学院生物研究所,河北,石家庄,050051 3. 河北农业大学生命科学学院,河北省农作物病虫害生物防治工程技术中心,河北,保定,071001 |
| 基金项目： | 现代农业产业技术体系建设专项资金资助 |
| 摘 要： | 利用Qiagen试剂盒提取花生害虫华北大黑鳃金龟幼虫蛴螬中肠总RNA,并纯化mRNA,然后按照Stratagene试剂盒合成双链cDNA,经过凝胶柱层析,收集符合要求的cDNA片段,加工后与Uni-ZAP XR Vector连接,经Gigapack Ⅲ Gold体外包装,即获得蛴螬的cDNA文库,未扩增文库滴度1.95×106pfu/mL,重组率为99.7%,扩增后文库滴度达1.34×109pfu/mL,插入cDNA片段的长度平均为1.34kb,表明成功构建了高质量的蛴螬中肠cDNA文库。 |
| 关 键 词： | 花生 蛴螬 中肠组织 cDNA文库 |
Construction of a cDNA Expression Library of Holotrichia oblita Larvae Midgut |
|
| LIU Xiao-min,ZHANG Xia,LI Jie,GUO Wei.Construction of a cDNA Expression Library of Holotrichia oblita Larvae Midgut[J].Journal of Peanut Science,2010,39(1):15-19. | |
| Authors: | LIU Xiao-min ZHANG Xia LI Jie GUO Wei |
| Institution: | 1.College of Plant Protection/a>;Agri.Uni.of Hebei/a>;Baoding 071001/a>;China/a>;2.Biological Institute of Hebei/a>;Shijiazhuang 050051/a>;3.College of Life Science/a>;Agri.Uni.of Hebei/Biological Control Centre of Plant Diseases and Pests of Hebei/a>;China |
| Abstract: | Midgut mRNA of Holotrichia oblita （H. oblita） larvae was isolated using the RNeasy Mini Kit and Oligotex mRNA purification Kit （Qiagen Inc. ）. A cDNA library was constructed from H. oblita midgut mRNA using the ZAP-cDNA Gigapack Cloning Kit （Stratagene）. eDNA was unidi- rectionatly ligated into the Uni-ZAP XR vector （Stratagene） between the EcoR I and Xho I sites and packaged with the Gigapack Ⅲ Gold package extract. The library has a complexity of 1. 95 × 10^6 plaques, of which over 99. 7% were recombinants. The titerring of amplified library was 1.34 × 10^9 pfu/mL. The average length of the insert cDNA fragments was over 1340 bps. It suggested that one high quality cDNA library of the H. oblita larvae midgut had been constructed. |
| Keywords: | peanut Holotrichia oblita midgut cDNA library |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |