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Virology ; 362(2): 283-93, 2007 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-17270231

RESUMO

Mouse cells are non-permissive to human immunodeficiency virus type 1 (HIV) in that there is a pronounced post-integration block to viral replication. We have recently demonstrated that mouse-human somatic cell hybrids that contain human chromosome 2 increase both HIV Capsid (CA) production and infectious virus release. Here we report on the isolation of three mouse-human microcell hybrids (MCHs) that behave similarly, starting from a pool of 500 MCH clones. Release of virus was specific to HIV and cell revertants that no longer contained any human chromosome fragments did not release CA or infectious virus. Two of the three cell clones were identical as judged by PCR STS content and fluorescence in situ hybridization (FISH) and contained a single 2-12 human chromosome chimera. The third cell clone only contained human chromosome 12, as determined by PCR, FISH, and microarray analyses. There were no consistent differences in Gag protein and spliced/unspliced viral RNA levels between mouse cell lines. CMV promoter-driven, codon-optimized gag-pol had no effect on infectious HIV release from these mouse cells, despite allowing Gag targeting and increasing CA production. These permissive mouse-human MCHs and their corresponding non-permissive revertants may prove useful for mechanistic studies and also for identifying the responsible gene(s) or factor(s) involved in the production of HIV.


Assuntos
HIV-1/crescimento & desenvolvimento , Células Híbridas/virologia , Animais , Linhagem Celular , Linhagem Celular Tumoral , Cromossomos Humanos Par 12 , Cromossomos Humanos Par 2 , Códon , DNA Viral/análise , Produtos do Gene gag/análise , Produtos do Gene gag/biossíntese , Proteína do Núcleo p24 do HIV/biossíntese , Humanos , Hibridização in Situ Fluorescente , Camundongos , Análise em Microsséries , Reação em Cadeia da Polimerase , Precursores de Proteínas/análise , RNA Viral/análise , RNA Viral/genética , Produtos do Gene gag do Vírus da Imunodeficiência Humana
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