Design and preparation of Epstein-Barr virus genome-wide cDNA probes
10.3321/j.issn:1673-4254.2005.03.002
- VernacularTitle:EBV病毒全基因组cDNA探针的设计与制备
- Author:
Wei-Yi FANG
1
;
Wen-Ling ZHENG
;
Wen-Li MA
;
Teng-Fei LIU
;
Shuang WANG
;
Wei-Bing XIE
;
Hong LI
;
Cai-Peng REN
;
Kai-Tai YAO
Author Information
1. 中南大学
- Keywords:
EBV genes;
EBV chip;
cDNA probes design;
gene cloning;
open reading frames
- From:
Journal of Southern Medical University
2005;25(3):246-250
- CountryChina
- Language:Chinese
-
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.
