一种新的土拨鼠α干扰素基因分型方法的建立及其意义  

A novel method for genotyping of woodchuck interferon alpha and its applications

作　　者：卢银平[1] 徐飏[2] 王宝菊[3] 董继华[1] 陆蒙吉[2] 杨东亮[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院病毒研究室,武汉430030 [2]德国埃森大学病毒学研究所 [3]华中科技大学同济医学院附属同济医院临床免疫研究室,武汉430030

出　　处：《中国免疫学杂志》2007年第6期551-554,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(30271170)

摘　　要：目的:建立新的土拨鼠α干扰素(IFN-α)基因分型方法,分析急性和慢性土拨鼠肝炎病毒(Woodchuck hepati-tis virus,WHV)感染的土拨鼠肝细胞中不同型别的IFN-α基因表达谱及其意义。方法:利用已知序列和基因型的土拨鼠IFN-α基因克隆质粒,建立基于土拨鼠IFN-α基因序列的限制性片段长度多态性的土拨鼠IFN-α基因分型方法。PolyIC体外刺激正常土拨鼠外周血单个核细胞(PBMC),调查正常土拨鼠PBMC受PolyIC刺激后IFN-α基因的表达谱。提取急性和慢性土拨鼠肝炎病毒感染的土拨鼠肝组织mRNA,RT-PCR扩增土拨鼠IFN-α基因,分子克隆后调查急性和慢性感染的土拨鼠肝细胞中IFN-α基因表达谱。结果:建立了土拨鼠IFN-α基因限制性片段长度多态性分型方法。急性和慢性感染的土拨鼠肝细胞中IFN-α基因的表达谱明显不同于正常动物的表达谱,假基因表达所占比例大。结论:新建立的土拨鼠IFN-α基因分型方法可用于分析土拨鼠不同组织中IFN-α的基因表达谱。Objective ：To establish a novel method of genotyping for woodchuck（ Marmota monax） interferon alpha（IFN-ot） to investigate the profdes of IFN-ot expressed in the liver of woodchuck with acute and chronic woodchuck hepatitis virus（WHV） infection. Methods： Analysing sequence data about different genotype woodchuck IFN-α and establishing a novel genotyping method for woodchuck IFN-α based on the restriction length polymorphism analysis. After drawing the total RNA of liver tissue with either acute or chronic WHV-infected, woodchuck IFN-α gene were amplified by RT-PCR and cloned. The profiles of woodchuck IFN-α gene expressed in the liver were analyzed. Naive woodchuck PBMCs were stimulated with high concentration PolyIC, and the expressing profiles of IFN--α were investigated. Results： A novel genotyping method for wIFN-α based on the restriction length polymorphism analysis was developed. Expression profiles of IFN-α in livers of woodchucks were changed after the animals were infected by WHV, and the pseudogenes made their majoricy. There was difference between expressing profiles of IFN-α in naive woodchuck liver and PBMC. Conclusion：The novel genotyping method for woodchuck IFN-α can be used to investigate the profiles of IFN-α expressed in different tissue of woodchuck infected by WHV. It will help for studying the action of different subtype IFN-α and the mechanism of antiviral of interferon.

关 键 词：土拨鼠 Α干扰素 基因分型 

分 类 号：R392.11[医药卫生—免疫学] R512.62[医药卫生—基础医学]