Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter
Identifieur interne : 001D17 ( Istex/Curation ); précédent : 001D16; suivant : 001D18Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter
Auteurs : Takekoshi Masataka [Japon] ; Maeda-Takekoshi Fumiko [Japon] ; Ihara Seiji [Japon] ; Sakuma Sadatoshi ; Watanabe Yasushi [Japon]Source :
- Gene [ 0378-1119 ] ; 1991.
English descriptors
- Teeft :
- Bgal, Bgal activity, Blue plaques, Chimeric, Chimeric gene, Chimeric iucz gene, Cytomegalovirus, Early stage, Foreign gene, Foreign genes, Fragment, Gene, Genome, Genome structures, Hcmv, Hcmv genome, Hcmvlacr, Iacz gene, Insertion, Iucz, Iucz gene, Late glycoprotein, Late stages, Marker genes, Mocarski, Negative regulation, Open circles, Opposite directions, Plasmid, Progeny genome, Promoter, Recombinant, Recombinant genomes, Recombinant virus, Recombinant viruses, Reversion rate, Simian virus, Viral, Viral proteins, Virion dnas, Virus replication, Virus vector.
Abstract
Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.
Url:
DOI: 10.1016/0378-1119(91)90413-6
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Sakuma Sadatoshi<affiliation><mods:affiliation>Molecular Oncology Division, Meiji Institute of Health Science, 540 Naruda, Odawara 250 Japan Tel. (0465)37-3661</mods:affiliation>
<wicri:noCountry code="subField">(0465)37-3661</wicri:noCountry>
</affiliation>
Le document en format XML
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<term>Chimeric iucz gene</term>
<term>Cytomegalovirus</term>
<term>Early stage</term>
<term>Foreign gene</term>
<term>Foreign genes</term>
<term>Fragment</term>
<term>Gene</term>
<term>Genome</term>
<term>Genome structures</term>
<term>Hcmv</term>
<term>Hcmv genome</term>
<term>Hcmvlacr</term>
<term>Iacz gene</term>
<term>Insertion</term>
<term>Iucz</term>
<term>Iucz gene</term>
<term>Late glycoprotein</term>
<term>Late stages</term>
<term>Marker genes</term>
<term>Mocarski</term>
<term>Negative regulation</term>
<term>Open circles</term>
<term>Opposite directions</term>
<term>Plasmid</term>
<term>Progeny genome</term>
<term>Promoter</term>
<term>Recombinant</term>
<term>Recombinant genomes</term>
<term>Recombinant virus</term>
<term>Recombinant viruses</term>
<term>Reversion rate</term>
<term>Simian virus</term>
<term>Viral</term>
<term>Viral proteins</term>
<term>Virion dnas</term>
<term>Virus replication</term>
<term>Virus vector</term>
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<front><div type="abstract" xml:lang="en">Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.</div>
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