现代食品科技2017，Vol.33Issue(3)：93-98,80,7.DOI:10.13982/j.mfst.1673-9078.2017.3.015
四种基因定量方法对实时荧光PCR与微滴式数字PCR检测霍乱弧菌基因表达量的分析
Real-time and Droplet Digital PCR Analyses for Detection of Vibrio cholerae Gene Expression by Four Gene Quantification Methods
摘要
Abstract
Gene expression assays are highly important for analyzing gene function and investigating biological mechanisms.Currently,real-time and droplet digital PCR are effective methods for measuring gene expression.sfs,vcc,RecA,hly,and 16S rRNA genes of Vibrio cholerae were used for experiments after low temperature treatment.The samples were assessed by real-time and droplet digital PCR separately before and after low temperature treatment.Real-time fluorescence PCR experimental data were analyzed using the AACT and geNorm methods.Droplet digital PCR experimental data were analyzed using the relative and absolute quantification methods to calculate the fold change in gene expression in the treatment group as compared to the control group.The data from the four gene quantification methods were analyzed using the multidimensional scaling method.Under these experimental conditions,the results indicated that there was a significant difference between the results of the four gene quantification methods.The expression level of the 16S rRNA gene changed,making it unsuitable as a reference gene.Droplet digital PCR could give more intuitive results regarding the amount of change in gene expression and have better accuracy in the relative quantitative analysis of differentially expressed genes.关键词
基因表达/实时荧光定量PCR/微滴式数字PCRKey words
gene expression/real-time PCR/droplet digital PCR引用本文复制引用
唐静,贾俊涛,赵丽青,王静,张健,姜英辉,徐彪,王昌军,马云..四种基因定量方法对实时荧光PCR与微滴式数字PCR检测霍乱弧菌基因表达量的分析[J].现代食品科技,2017,33(3):93-98,80,7.基金项目
国家质检总局科研计划(2012IK305,2013IK175,2014IK114) （2012IK305,2013IK175,2014IK114）
