摘要
[目的]构建水稻osvdac3和osvdac5基因RNA干涉表达载体,获得转干涉载体的转基因水稻植株。[方法]采用TRIzol法提取水稻幼苗总RNA,以反转录的cDNA为模板,扩增得到osvdac3和osvdac5的分别靠近5’端和3’端约500～600bp的特异序列,用Gateway重组克隆技术构建RNA干涉表达载体,采用农杆菌介导的愈伤组织侵染法进行遗传转化。[结果]成功构建水稻2个osvdac3和2个os-vdac5基因RNA干涉表达载体,并获得转osvdac3干涉载体的阳性植株。[结论]干涉osvdac3和osvdac5的植株为研究其功能提供材料。
[Objective] The research aimed to construct the RNAi expression vectors of osvdac3 and osvdac5 gene,and to obtain transgenic plants with the RNAi vectors.[Method] The total RNA was extracted from rice seedlings with TRIzol and reverse transcripted to cDNA.4 special fragments about 500-600 bp nearby 5’ and 3’ of osvdac3 and osvdac5 gene were amplified with the cDNA template.The gateway recombination cloning technology was used to construct the RNAi exprefssion vector for 4 fragments.The recombinant RNAi expression vector plasmids were transformed into the callus through agrobacterium-mediated approach.[Result] 4 RNAi expression vectors of the osvdac3 and osvdac5 gene were constructed successfully.And these RNAi expression vectors were introduced into rice variety Yuetai B,and some transgenic plants were obtained.[Conclusion]The transgenic plants with RNAi for osvdac3 and osvdac 5 provided the matercal for studying their function.
出处
《安徽农业科学》
CAS
北大核心
2011年第28期17176-17178,共3页
Journal of Anhui Agricultural Sciences
基金
国家自然基金项目(30871318)
中南民族大学科研基金项目(YZY07007)
