第一军医大学学报2005，Vol.25Issue(3)：246-250,5.
EBV病毒全基因组cDNA探针的设计与制备
Design and preparation of Epstein-Barr virus genome-wide cDNA probes
摘要
Abstract
Objective To design and clone all known and predicted coding genes of Epstein-Barr virus (EBV) as the cDNA probes for preparing the microarray for EBV detection, thereby to facilitate further investigation of the pathogenetic role of EBV. Methods Oligo 6.0 software, BLAST program and Primer Premier 5 software were employed to design and screen the cDNA probes of the whole EBV genome, whose length ranged from 300 to 600 mer each with high specificity. These cDNA probes obtained through PCR and reverse transcriptase (RT)-PCR amplification from the genomic DNA and RNA of B95-8 cells and nasopharyngeal carcinoma (NPC) tissue were cloned into T/A clone vector, followed by identification of these probes by sequencing analysis. Result and Conclusion A total of 85 gene fragments (BWRF1 genecontained 7 repeats of open reading frames) coding for proteins and 2 EBERs in EBV genome were successfully cloned, not including LF1 and LF3 genes that did not exist in EBV genome ofB95-8 cells, which provides the basis for preparing microarray to explore the role of EBV genome in its related diseases.关键词
EBV基因/cDNA探针/EBV基因芯片/基因克隆Key words
EBV genes/EBV chip/cDNA probes design/gene cloning/open reading frames分类
医药卫生引用本文复制引用
方唯意,郑文岭,马文丽,刘腾飞,王爽,谢卫兵,李虹,任彩萍,姚开泰..EBV病毒全基因组cDNA探针的设计与制备[J].第一军医大学学报,2005,25(3):246-250,5.基金项目
This study is sponsored by Guangzhou Science and Technology Commission as a key research project (2004z2-e0111) （2004z2-e0111）
