雏鸭肝炎病毒侵染下肝脏消减cDNA文库的构建及差异基因筛选

本研究通过构建雏鸭肝脏消减cDNA文库,旨在筛选并鉴定与雏鸭病毒性肝炎相关的基因,对相关基因进行功能聚类分析进而探究其作用机理。利用抑制性消减杂交(Suppression subtraction hybridization,SSH)技术构建3日龄健康全同胞金定鸭人工感染雏鸭肝炎病毒(Duck hepatitis virus,DHV)与同期注射等量生理盐水差异表达基因的SSH-cDNA文库。对其中563个阳性克隆进行测序,共获得299条差异表达序列标签(Expres sedsequence tags,ESTs)。去除冗余的cDNA序列载体并聚类拼接后,进行核酸和蛋白质同源性的比较和功能聚类分析。结果表明:有70个不同的基因与ESTs具有高度的同源性(E值150bp,匹配度>80%),且多数基因与细胞组分合成、信号转导以及病理状态下的生物学调控过程相关。I型雏鸭病毒性肝炎的发生和发展是多基因多步骤的复杂过程,该结果为深入研究雏鸭肝炎病的分子调控机制提供基础。

Construction of a Suppression Subtractive Hybridization cDNA Library to Screen Differentially Expressed Genes from Duck Liver Infected Duck Hepatitis Virus

This research aimed to detect and identify genes associated with duck viral hepatitis(DVH) by construction of suppression subtractive cDNA library,and explore their mechanism by function cluster analysis.Using suppression subtraction hybridization(SSH),a different expression SSH-cDNA library of 3-day-old and full-sib ducklings infected artificially by DHV was constructed and the same day-age ducklings were injected with the same quantity of saline.Of which 563 clones were sequenced,and 299 differentially expressed sequence tags(ESTs) were obtained.After dislodging the redundant cDNA sequence and clustering splicing,the software BLAST in NCBI of GenBank was used to do the nucleic acid and protein homology comparisons and functional analysis.The results showed that 70 different genes had highly homologous with ESTs(E value 150 bp,match rate>80%),most of which were related to the synthesis of cell components,signal transduction and the biological control process of pathological conditions.The occurrence and development of DVH was a complex multistep process involving multiple genes.The results provide a basis for further study of the molecular mechanism of DVH.