热带作物学报2011，Vol.32Issue(10)：1833-1837,5.DOI:10.3969/j.issn.1000-2561.2011.10.009
农杆菌浸润瞬时表达分析番木瓜miR162a的研究
Transient Assays of Papaya miR162aProcessing by Agro-infiltration
摘要
Abstract
The most widely used method for small RNA analysis in plants is agro-infiltration,which makes use of Agrobacterium tumefaciens to deliver transgenes into leaf cells of Nicotiana benthamiana Compared to the traditional method of stably transformed plants,agro-infiltration is more rapid,and samples can be analyzed just a few days after inoculation. Here,a method was described for analysis of papaya mir162a processing using agro-infiltration of N.benthamiana leaves.Cpa-mir162a precursor was amplified by PCR from papaya genomic DNA and ligased to pSuper1300 generating the final binary plasmid and then delivered to Agrobacterium tumefaciens strain GV3101 for agro-infiltration of N.benthamiana leaves.RNA was analyzed by Northern blot after 48 h inoculation,mature miR162a was detected,but not from wild type N.benthamiana leaves and papaya leaves because the low abundance of mir162a.This ago-infiltration method would help to the structure-function analysis and processing mechanism of papaya microRNAs in vitro.关键词
番木瓜/瞬时表达分析/农杆菌浸润/microRNAKey words
Papaya/transient assay/Agroinfiltration/microRNAs分类
生物科学引用本文复制引用
彭军,杨腊英,王国芬,郭立佳,汪军,黄俊生..农杆菌浸润瞬时表达分析番木瓜miR162a的研究[J].热带作物学报,2011,32(10):1833-1837,5.基金项目
国家科技支撑计划(2008BD02),农业部行业科技项目(200903049),中国热带农业科学院环境与植物保护研究所中央级公益性科研院所基本科研业务费(2009hzslJ007). （2008BD02）
