Design and preparation of Epstein-Barr virus genome-wide cDNA probes.
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.


CONCLUSIONS
A total of 85 gene fragments (BWRF1 gene-contained 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 of B95-8 cells, which provides the basis for preparing microarray to explore the role of EBV genome in its related diseases.
Fang
Wei-Yi
WY
Institute of Cancer Research, Xiangya Medical School, Central South University, Changsha 410078, China. fangweiyi1975@yahoo.com.cn
Zheng
Wen-Ling
WL
Ma
Wen-Li
WL
Liu
Teng-Fei
TF
Wang
Shuang
S
Xie
Wei-Bing
WB
Li
Hong
H
Ren
Cai-Peng
CP
Yao
Kai-Tai
KT
D001483:Base Sequence; D015342:DNA Probes / Q000096:biosynthesis* / Q000235:genetics; D018076:DNA, Complementary; D020031:Epstein-Barr Virus Infections / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods; D008969:Molecular Sequence Data; D009303:Nasopharyngeal Neoplasms / Q000821:virology; D016366:Open Reading Frames / Q000235:genetics
D015342:DNA Probes; D018076:DNA, Complementary