摘要
为研究水孔蛋白基因EgPIP1 3在洋桔梗(Eustoma grandiforum)采后切花中的表达特点,采用肌动蛋白(β-actin)基因为内参,以洋桔梗花瓣cDNA为模板,通过对PCR体系循环数及退火温度的优化,建立了一个稳定、特异的半定量反转录PCR(Reverse transcription PCR,RT-PCR)体系.运用此体系检测了EgPIP1 3在洋桔梗切花中的表达.结果表明,EgPIP1 3基因在洋桔梗切花的茎、叶、萼、花瓣、雄蕊和雌蕊中均有表达,但表达量存在明显的差异,依次表现为茎>雌蕊>萼>叶>雄蕊>花瓣.
A stable and specific semi-quantitative reverse transcription PCR （RT-PCR） system for the expression analysis of EgPIP1;3 gene in different organs of cut eustoma （Eustoma grandiforum） flowers was established, using β-actin gene as the internal control, and the eDNA from eustoma petal as the template. Cycle numbers and annealing temperature for this system were optimized. Based on the sys- tem, the expression of EgPIP1; 3 gene in different tissues of cut eustoma flowers was analyzed. The re- suits showed that the gene could be expressed in all detected tissues of cut eustoma flowers, including stem, leaf, calyx, petal, stamen and pistil. However, there were significant differences in mRNA lev- els among them. The highest expression level was detected in stems, then immediately followed by pis- tils, while moderate expression levels were in calyxes, leaves and stamina, and the lowest level was in petals.
出处
《仲恺农业工程学院学报》
2012年第1期6-10,共5页
Journal of Zhongkai University of Agriculture and Engineering
基金
国家自然科学基金(3107182930771519)
广东省自然科学基金(10151022501000035
8251022501000002)
仲恺农业工程学院研究生创新基金(H1410010)资助项目
