河南农业科学2013，Vol.42Issue(3)：46-49,4.
利用烟草的cDNA酵母表达文库筛选镉胁迫相关基因
Isolation of Cadmium Stress Related Genes in Tobacco with cDNA Expression Library in Yeast Mutant
摘要
Abstract
To facilitate the identification of functional genes in tobacco under heavy metal stress, this study constructed a cDNA expression library with Gateway technology. The tobacco seedlings were treated with different heavy metal solutions,and then the roots were harvested and the total RNAs were extracted. After mRNAs were isolated,the reverse transcription assay was performed with SMART technology. Then the ds-cDNAs were cloned into pDONR222 vector and LR reaction was performed to transfer cDNA into yeast expression vector, pDEST22, finally forming the tobacco cDNA expression library. The average titer of this library was 4. 32×106 cfu/mL,and the quality of the destination cDNA library met the requirements of following experiments. The cadmium stress-related genes of tobacco were screened by using cadmium-sensitive yeast mutation strain Aycfl ,and more than 70 clones were obtained, which involved 24 different genes such as the genes coding glutathione S-transferase, copper chaperone and metallothionein. All of these genes were considered as the candidate genes of heavy metal stress-related genes in tobacco.关键词
烟草/重金属/镉胁迫/cDNA文库/功能基因Key words
tobacco/ heavy metal/ cadmium stress/ cDNA library/ functional genes分类
生物科学引用本文复制引用
李静,王晶,张美..利用烟草的cDNA酵母表达文库筛选镉胁迫相关基因[J].河南农业科学,2013,42(3):46-49,4.基金项目
中国科学院植物资源保护与可持续利用重点实验室基金资助项目(201220zs) （201220zs）
