摘要
合适的内参基因是采用实时荧光定量PCR研究基因表达获得可信数据的基础.为了获得合适的内参基因,更好地研究烟草花叶病毒(Tobacco mosaic virus,TMV)和马铃薯X病毒(Potato virus X,PVX)与烟草和番茄2种寄主的互作,本项研究根据已报道的云烟和番茄看家基因设计引物,使用geNormPlus软件分别对其在TMV和PVX侵染下的云烟97和番茄中表达量的稳定性进行了评估.结果表明:2种寄主植物中供试看家基因中各有4对引物满足qPCR的要求,分别是云烟tub,CYP,GAPDH,β-tub,番茄18S,UK,ACT,EFl-α.在2种病毒侵染后的云烟内参基因中表达最稳定的为CYP,番茄中内参基因为ACT.最终确定这2个看家基因分别作为实时荧光定量PCR研究烟草和番茄在TMV和PVX胁迫下寄主体内基因表达变化的内参基因.该结果为进一步探讨2种病毒共同侵染时,它们与烟草和番茄2种寄主互作生物学效应研究奠定了基础.
It is the base to select proper reference genes for getting valid data to analyze gene expression by quantitative real-time PCR（RT-qPCR）.In order to get proper reference genes of tobacco and tomato infected by TMV or PVX for studying the interaction between the viruses and their hosts,we designed the primer pairs of the housekeeping genes of tobacco and tomato based on the previous studies,and evaluated their expression stability in TMV or PVX infected tobacco and tomato with geNormPlus software.The results showed that the primer pairs of tub,CYP,GAPDH andβ-tub in tobacco and the primer pairs of18 S,UK,ACT and EFl-αin tomato matched the request of RT-qPCR.CYP was the most stable housekeeping gene in tobacco infected by TMV or PVX,and ACT was the most stable housekeeping gene in tomato.So,these two genes were determined to be the reference genes normalization of qRT-PCR data in tobacco or tomato in virus-host interaction system,which provides a base for studying the biological effect of the interaction between TMV or PVX and its hosts.
出处
《西南大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第7期13-18,共6页
Journal of Southwest University(Natural Science Edition)
基金
国家自然科学基金(30900937)
重庆市自然科学基金(CSTC2010BB1122)
中央高校基本科研业务费(XNJI2009B023
2362014xk08)
