【摘要】 提取经重金属胁迫处理的水稻幼苗总RNA并纯化mRNA,采用SMART技术反转录成cDNA,利用Gateway技术将cDNA构建到入门载体pDONR222后,再经LR反应重组到酵母表达载体pYES2-DEST52上,获得水稻cDNA表达文库。检测表明,所构建的cDNA文库能达到后续试验要求。使用重金属镉敏感酵母突变株ycf1Δ进行水稻中重金属镉耐受相关基因筛选,获得多个可以恢复表型的酵母单克隆。提取质粒并测序分析后可知涉及到水通道蛋白、蛋白酶抑制剂以及金属硫蛋白等不同基因,筛选所得基因即为水稻应答重金属镉胁迫的候选基因。更多还原

【Abstract】 Total RNAs of rice which treated by heavy metal were isolated and purified,then the ds-cDNA were reversely transcripted and were cloned into pDONR222 and pYES2-DEST52 through BP and LR reaction separately,formed the rice cDNA express library.Then the library was introduced into yeast mutant ycf1Δ and several yeast clones were obtained on YNB culture medium plate.The plasmids in these clones were sequenced and different cDNAs were isolated,including aquaporin,Bowman Birk trypsin inhibitor,metallothionein,and so on.更多还原