摘要
[目的]克隆线粒体相关基因nad1,获得转nad1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cD-NA为模板,扩增得到nad1;将nad1接到线粒体信号肽Rf1b的5’(Rf1b5’),装载到pCAMBIA1305.1双元表达载体,采用农杆菌介导的愈伤组织侵染法进行遗传转化。[结果]克隆的目的基因nad1大小为978bp,成功构建了携带线粒体信号肽的nad1植物表达载体,并获得了转nad1基因的阳性植株。[结论]为探讨水稻中过表达nad1对水稻生长的影响奠定了基础。
[Objective] The aim was to clone the mitochondria related gene nad1 and obtain transgenic rice plants with nad1.[Method] The total RNA was extracted from rice seedlings and reverse transcriped to cDNA.The target gene nad1 was amplified with the cDNA as template.nad1 and Rf1b,a sequence of signal peptide of mitochondria,were linked to binary expression vector pCAMBIA1305.1.The recombinant plasmid was transformed into the callus by Agrobacterium-mediated approach.[Result] The target gene nad1 was 978 bp.The binary expression vector carried nad1 and signal peptide of mitochondria was constructed successfully.A lot of transgenic plants were obtained.[Conclusion] The study will provide basis to investigate the effect of over-expression of nad1 on rice plant growth.
出处
《安徽农业科学》
CAS
北大核心
2011年第28期17158-17160,共3页
Journal of Anhui Agricultural Sciences
基金
中南民族大学自然基金重点项目(YZZ08010)
