欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (18): 189-193.doi: 10.11924/j.issn.1000-6850.2011-3270

所属专题： 生物技术； 马铃薯； 烟草种植与生产

• 生物技术科学 • 上一篇    下一篇

烟草马铃薯Y病毒病相关基因eIF4E的片段克隆及RNAi载体构建

皮金鹏   

  • 收稿日期:2011-11-07 修回日期:2012-01-23 出版日期:2012-06-25 发布日期:2012-06-25
  • 基金资助:

    云南省烟草专卖局（公司）科技项目

Cloning of PVY associated gene eIF4E and construction of its corresponding RNAi vectors

  • Received:2011-11-07 Revised:2012-01-23 Online:2012-06-25 Published:2012-06-25

PDF (PC)

1226

摘要：

以感病烟草品种为材料，提取总RNA，根据已报道的eIF4E基因cDNA的序列，在保守区域和差异区域分别设计特异性引物，采用RT-PCR克隆出eIF4E基因的目的片段。以中间载体pKANNIBAL和表达载体pART27为基础，构建成以CaMV35S启动子为驱动的含有“正向eIF4E目的片段-pdk intron-反向eIF4E目的片段”的RNAi烟草表达载体，构建的RNAi载体将抑制eIF4E基因表达，为了解其基因功能和烟草抗病毒育种打下基础。

关键词: 耐受性, 耐受性

Abstract:

The total RNA was isolated from the PVY susceptible tobacco variety,the specific primers was designed according to the high homology and differential region of the published sequence of eIF4E gene,cloned the target gene fragment of eIF4E gene by RT-PCR. Then the promoter CaMV 35S driven,containing " Sense eIF4E gene target fragments-pdk intron-Antisence eIF4E gene target fragments",tobacco RNAi expression vector were constructed based on the intermediate vector pKANNIBAL and expression vector pART27. The RNAi plasmid is constructed to inhibit the expression of eIF4E gene,in order to understand the gene and lay the foundation for the breeding of tobacco viral resistance.