title: Characterization of late gene expression factor LEF-10 from Bombyx mori nucleopolyhedrovirus.
abstract: The LEF-10 expression factor from the Bombyx mori nuclear polyhedrosis virus (BmNPV) does not have significant homology with other late expression factors and is thought to be a transcriptional cofactor. To investigate the function of LEF-10, a Red recombination system was used to knock out the lef-10 gene from the BmNPV genome and a lef-10 gene knockout virus (ko-Bacmid) was constructed. The lef-10 gene was repaired back to the viral genome using a Bac-to-Bac system to create the repaired virus (re-Bacmid). When ko-Bacmid was transfected into BmN cells, the detected titer of progeny virus in the medium was zero, whereas the titer of the progeny re-Bacmid remained at a level similar to that of the wild type virus (wt-Bacmid). The viral DNA replication, transcription and expression of viral early, late and very late genes after ko-Bacmid transfection into BmN cells were evaluated. The quantitative polymerase chain reaction showed that the ko-Bacmid viral genome replication level remained low and that the ko-Bacmid viral gene transcription level was significantly lower than those of wt-Bacmid and re-Bacmid. No expression of the early gene lef-3 was detected. These results suggest that the lef-10 gene has significant effects on DNA replication of the viral genome and BmNPV gene transcription at each phase and deletion of the lef-10 gene affects the level of expression of the viral early gene directly.
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, Zhejiang Province, Hangzhou 310018, China. mikkyu@163.com
authors.forename: Wei
authors.initials: W
authors.lastname: Yu
authors.forename: Chao-Yi
authors.initials: CY
authors.lastname: Du
authors.forename: Yan-Ping
authors.initials: YP
authors.lastname: Quan
authors.forename: Zuo-Ming
authors.initials: ZM
authors.lastname: Nie
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Zheng-Bing
authors.initials: ZB
authors.lastname: Lv
authors.forename: Yao-Zhou
authors.initials: YZ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D012831:Bombyx / Q000821:virology; D004261:DNA Replication; D015967:Gene Expression Regulation, Viral; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014157:Transcription Factors; D014764:Viral Proteins