RESUMO
We used a series of deletion mutations in the 5' untranslated region of the prototype D type retrovirus, Mason-Pfizer Monkey Virus (MPMV), to analyse RNA encapsidation. A region was identified upstream of the major splice donor which reduced particle production but had a proportionally greater effect on RNA packaging. A small deletion downstream of the splice donor had little effect on RNA production and caused no significant packaging defect. A large deletion encompassing the end of the primer binding site down to the splice donor had a dramatic effect, disrupting viral protein synthesis. Stable cell lines were produced containing packaging-defective virus. These first-generation packaging cell lines were used to package and transfer an MPMV-based vector.
Assuntos
Regiões 5' não Traduzidas/genética , Vírus dos Macacos de Mason-Pfizer/fisiologia , RNA Viral/genética , Replicação Viral/genética , Regiões 5' não Traduzidas/química , Animais , Células COS , Capsídeo/genética , Capsídeo/metabolismo , Linhagem Celular , Chlorocebus aethiops , Humanos , Rim , Vírus dos Macacos de Mason-Pfizer/genética , Mutagênese , DNA Polimerase Dirigida por RNA/genética , DNA Polimerase Dirigida por RNA/metabolismo , Deleção de Sequência , Proteínas Virais/genética , Proteínas Virais/metabolismo , Vírion/genética , Vírion/fisiologiaAssuntos
Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Enalapril/uso terapêutico , Transplante de Rim , Policitemia/prevenção & controle , Complicações Pós-Operatórias , Adulto , Inibidores da Enzima Conversora de Angiotensina/efeitos adversos , Bloqueadores dos Canais de Cálcio/efeitos adversos , Bloqueadores dos Canais de Cálcio/uso terapêutico , Enalapril/efeitos adversos , Eritropoetina/sangue , Feminino , Hematócrito , Humanos , Masculino , Nifedipino/efeitos adversos , Nifedipino/uso terapêutico , Estudos Retrospectivos , Fatores de TempoRESUMO
Glycosylation of lipoprotein lipase (LPL) was studied in human subcutaneous lipomas. Heparin-releasable LPL activities were higher in lipomas than those in adjacent normal adipose tissues, and showed good correlation with cellular LPL protein mass. Molecular weight of LPL subunit was 57 kDa in both tissues. After endoglycosidase H-digestion, two types of LPL subunits were found in normal adipose tissues; partially sensitive (55 kDa) and totally sensitive (52 kDa) form. In lipoma tissues, the fraction of partially sensitive form (55 kDa) was increased comparing with control adipose tissues. These results suggest that partially sensitive subunits constitute the major secretable form of LPL in human subcutaneous lipomas.
