慢性乙型肝炎患者炎症相关差异表达基因cDNA文库的构建和筛选  

Construction and Analysis of cDNA Library Accociated with Inflammation of Chronic Hepatitis B

作　　者：唐翠兰[1] 彭国平[1] 陈智[1] 朱海红[1] 郑敏[1] 

机构地区：[1]浙江大学医学院附属第一医院传染病研究所,杭州310031

出　　处：《同济大学学报（医学版）》2006年第1期4-8,共5页Journal of Tongji University(Medical Science)

基　　金：浙江省科技厅重点资助项目(2003C13015)

摘　　要：目的筛选慢性乙型肝炎(chron ic hepatitis B,CHB)患者与无症状乙型肝炎病毒携带者(asymptom atic HBVcarriers,ASCs)表达差异的基因。方法分离CHB及ASCs的外周血单个核细胞(peripheral b lood mononuc lear cells,PBMC)并提取RNA,SMART技术合成cDNA,采用抑制消减杂交(suppression subtractive hybrid ization,SSH)技术构建差异表达基因cDNA文库,进一步采用dot b lot筛选,得到差异表达的基因。结果经过筛选共获得了22个阳性克隆,通过序列分析,发现它们代表22个基因,在CHB表达增高的基因多是和炎症相关,在ASCs表达增高的基因中,多为新基因。结论成功构建了CHB与ASCs的消减文库并初步分析这些差异表达基因的意义,对于阐明CHB的发病机制及其治疗都有重要意义。Objective To identify differentially expressed genes between chronic hepatitis B （CHB） patients and asymptomatie HBV carriers （ASCs）. Methods eDNA was synthesized by SMART technology from RNA of peripheral blood mononuclear cells （PBMC）, subtracted libraries were constructed by suppression subtractive hybridization （SSH） and rescreened by dot blot hybridization. Results 22 clones were identified as differentially expressed genes after screening, and they were homologous to 22 genes. The majority of genes upregulated in CHB was associated with inflammation, but most of the upregulated genes in ASCs were novel genes. Conclusion Subtracted libraries have constructed successfully and the biological sense of the genes were analyzed, which might provide clues in elucidating the mechanisms of the two different HBV infection states and designing therapeutic targets for hepatitis B infection.

关 键 词：慢性乙型肝炎 无症状乙型肝炎病毒携带者 抑制消减杂交 基因表达 

分 类 号：R512.62[医药卫生—内科学]