Use of a glycoprotein gB promoter for expression of genes inserted into the human cytomegalovirus genome.
We attempted to utilize the human cytomegalovirus (HCMV) as an expression vector by replacing the dispensable genes of the viral genome with foreign genes. The selection of a promoter to be fused to the foreign gene is important to achieve a high expression rate in the recombinant virus. We selected the glycoprotein B (gB) promoter of HCMV as a target of analysis because gB is one of the most abundantly synthesized components in cell culture. The gB promoter, fused to the E. coli lacZ gene, was introduced into the HCMV HindIII-O fragment region by homologous recombination. It was confirmed that the gB promoter-lacZ construct was inserted in the targeted site of HCMV. The expression of the lacZ gene in the recombinant virus infection was initiated 24 h after infection and increased until 120 h post infection. The lacZ gene expression was inhibited by the presence of cytosine arabinoside. These observations indicate that the expression of the lacZ gene is under the control of the late promoter of gB.
Takekoshi
M
M
Department of Molecular Life Science 2, School of Medicine, Tokai University, Isehara, Japan. mtakekos@is.icc.u-tokai.ac.jp
Maeda-Takekoshi
F
F
Ihara
S
S
Sakuma
S
S
Watanabe
Y
Y
D002460:Cell Line; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007763:Lac Operon; D011401:Promoter Regions, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*