摘要
硫氧还原蛋白基因OsTxnDC9是水稻miRNA3026的宿主基因。克隆出了水稻miRNA3026启动子,其总长度为1 477bp;构建出4个缺失片段,瞬时表达表明这个启动子为弱启动子。在此基础上克隆出水稻硫氧还原蛋白OsTxnDC9基因,其长度为480bp。生物信息学表明OsTxnDC9基因编码的氨基酸序列与二穗短柄草硫氧还原蛋白基因编码的氨基酸(XP_003573612.1)序列同源性最高。荧光定量PCR发现OsTxnDC9在水稻花粉一核中表达量最高,亚细胞定位表明其主要在细胞质中表达。这为探索miRNA3026启动子和宿主基因的关系奠定了基础。
The 1 477 bp promoter of miRNA3026 was cloned,furthermore,four mutant fragments with different length of deletion were constructed and transient expression assay showed that the activity of miRNA3026 promoter is weak.OsTxnDC9 was cloned,the host gene of miRNA3026,which has an open reading frame of482 bp.Bioinformatic analysis indicated that OsTxnDC9 encodes a protein which is highest homology with Brachypodium distachyon.The expression of OsTxnDC9 was highest in one nuclear stage of pollen,which was examined by real-time polymerase chain reaction essay.Subcellular localization suggested that the protein was located in cytoplasm.This research laied the foundation for further exploring the relationship of miRNA3026 promoter and its host gene.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2016年第1期29-37,共9页
China Biotechnology
基金
国家自然科学基金(31070276和31270360)
四川省国际合作基金(2012HH0006)资助项目
关键词
硫氧还原蛋白
启动子
宿主基因
瞬时表达
荧光定量
Thioredoxin
Promoter
Host gene
Transient expression
Real-time polymerase chain reaction
