基因结构分析软件对实时定量PCR引物进行设计和筛选  

Design and Screen of Primers in Real-time PCR using Gene Structure Analysis Software

作　　者：赵焕英[1] 赵君朋[1] 方霄[2] 杨颖[1] 张进禄[1] 

机构地区：[1]首都医科大学医学实验与测试中心,北京100069 [2]济宁医学院药学院,13级药物制剂技术山东日照276826

出　　处：《现代生物医学进展》2015年第36期7190-7193,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81200783);北京市财政支持基础临床合作基金项目(13JL28;14JL29);北京市财政支持校基金技术类项目(2014JS08)

摘　　要：目的:以人丝裂原活化蛋白激酶3(mitogen-activated protein kinase 3,MAPK3)基因结构为例,利用不同生物相关软件分析、设计和筛选合适的定量PCR引物。方法:利用NCBI的Gene数据库查找人MAPK3基因的参考序列、Uni Gene数据库查找标准参考序列;并用在线软件如Spidey,UCSC,Ensembl等分析基因结构;利用Primer3,Oligo6,IDT等软件进行引物设计;用MFOLD程序分析基因二级结构后,选择引物可定位的外显子位置;利用电子PCR进行引物扩增特异性的检验;最后通过实验检验引物的扩增效果。结果:从程序软件推荐的引物列表中筛选出一对能特异扩增人MAPK3基因的引物。结论:基因结构分析软件有助于定量PCR引物的设计。Objective： To analyze, design and screen the primers for mitogen-activated protein kinase 3（MAPK3） gene amplified in real-time quantitative PCR（q PCR） by using different biological softwares. Methods： Searching the sequence of Homo MAPK3 gene in NCBI gene database; searching standard sequence in Unigene database; analyzing the gene structure with online software such as Spidey,UCSC and Ensembl; designing the primers with Primer3, Oligo6 and IDT softwares; Localizing the primers in exon following analysis of secondary structure of gene with MFOLD program; testing the specificity of amplified products with Electronic PCR（e PCR）; testing the efficiency of amplification by experiment. Results： A pair of primers which can be used to specifically amplify MAPK3 gene were screened from the primer list proposed by software. Conclusions： The softwares for analysis of gene structure are helpful for designing primers used in q PCR.

关 键 词：基因结构 引物 实时定量PCR 

分 类 号：Q81[生物学—生物工程]