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鸡β-actin基因实时荧光定量PCR方法的建立
引用本文:马莉,谢秀兰,岳华.鸡β-actin基因实时荧光定量PCR方法的建立[J].中国畜牧兽医,2007,34(2):73-75.
作者姓名:马莉  谢秀兰  岳华
作者单位:西南民族大学生命科学与技术学院,成都 610041
摘    要:根据GenBank上鸡β-actin基因的序列,在保守区域设计并合成一对引物,采用SYBR Green I染料建立了荧光定量PCR法(real-time PCR)。以常规PCR产物为标准品,建立了标准曲线,并进行了熔解曲线分析。结果表明,标准曲线Ct值检测范围为12~31,扩增效率为95.1%;熔解曲线分析结果显示产物特异的单个峰,其Tm为88±0℃,检测周期从RNA提取到荧光定量PCR结束只需4 h。本试验建立的鸡β-actin基因实时荧光定量PCR法扩增效率高、检测范围广、检测周期短,为β-actin基因作为内参基因进行鸡功能基因与病原基因表达的定量分析奠定了基础。

关 键 词:  β-actin基因  实时荧光定量PCR  内参基因    
文章编号:1671-7236(2007)02-0073-03
修稿时间:2006-01-06

Establishment of Real- time PCR for β-actin Gene of Chicken
MA Li,XIE Xiu-lan,YUE Hua.Establishment of Real- time PCR for β-actin Gene of Chicken[J].China Animal Husbandry & Veterinary Medicine,2007,34(2):73-75.
Authors:MA Li  XIE Xiu-lan  YUE Hua
Affiliation:College of Life Science and Technology,Southwest University for Nationalities, Chengdu 610041, China
Abstract:According to the chicken β-actin gene sequences available in GenBank, a pair of primers was designed for establish a SYBR Green I quantitative real-time PCR method for β-actin gene of chicken. To establish the standard curve, the product of conventional PCR served as a standard. The analysis of melting curve was also carried out. The results show the linear range of Ct value was from 12 to 31 with a good correlation coefficient(r=0.996). The melting curve show a single peak with a 88±0℃ Tm value. The real-time PCR assay developed in this study can detect β-actin in expand range with high efficiency and less time. The assay provide the basis for β-actin gene of chicken as a reference gene in quantitative analysis of mRNA expression.
Keywords:chicken  β-actin gene  real-time PCR  reference gene
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