title: A comprehensive library of mutations of Epstein Barr virus.
abstract: A mutant library of 249 mutants with mutations that span the entire Epstein-Barr virus (EBV) genome was generated by transposition with EZ : : TN <KAN-2> and insertion with an apramycin resistance gene by a PCR-targeting method. This study also demonstrates the feasibility of generating deletions and site-specific mutations in the BRLF1 promoter on the EBV genome to determine the regions in the promoter that are crucial to transcription. Analysing BZLF1 and BRLF1 mutants by microarray analysis revealed that these two genes regulate the transcription of EBV lytic genes differently. A BZLF1 mutation affects global expression of EBV lytic genes; almost no lytic gene is expressed by the mutant after lytic induction. However, although a BRLF1 mutant still transcribes most lytic genes, the expression of these lytic genes is inefficient. Furthermore, this study shows that the proximal Zta-response element in the BRLF1 promoter is crucial to BRLF1 transcription from the EBV genome, despite the fact that another work demonstrated that this site was unimportant in transient transfection analysis. Furthermore, mutants with a mutation in BDLF1 and BORF1 cannot assemble viral capsids. Results of this study demonstrate the usefulness of a comprehensive mutant library in genetic analyses of EBV.
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Ya-Fang
authors: YF
authors: Chiu
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Chao-Ping
authors: CP
authors: Tung
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Yu-Hisu
authors: YH
authors: Lee
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Wen-Hung
authors: WH
authors: Wang
authors: Department of Applied Microbiology, National Chiayi University, Chiayi City 600, Taiwan.
authors: Ching
authors: C
authors: Li
authors: Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung 402, Taiwan.
authors: Jia-Yan
authors: JY
authors: Hung
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Chen-Yu
authors: CY
authors: Wang
authors: Division of Viral Infection, Department of Infectious Disease Control, International Research Center for Infections Diseases, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-Ku, Tokyo 108-8639, Japan.
authors: Yasushi
authors: Y
authors: Kawaguchi
authors: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang Gung University, Taoyuan 333, Taiwan.
authors: Shih-Tung
authors: ST
authors: Liu
mesh_terms: D004279:DNA, Viral / Q000235:genetics; D024882:Drug Resistance, Viral / Q000235:genetics; D015723:Gene Library; D016679:Genome, Viral; D015698:Genomic Library; D004854:Herpesvirus 4, Human / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics
chemical_list: D004279:DNA, Viral; D008562:Membrane Glycoproteins; D014764:Viral Proteins