RESUMEN
The DNA coding for the fusion protein of thromobopoietin mimetic peptide (TMP) and human IgG1 Fc fragment was amplified from recombinant plasmid pET28a/TMPFc, inserted into pPICZalphaA and transformed into Pichia pastoris using electroporation. The recombinants of correct phenotype were identified after screening on MDH and MMH culture medium. The fusion gene was verified with PCR and western blot. MTT method was used to test the activity of TMPFc in promoting the growth of Ba/ F3-mpl cell. The TMPFc with a 64 000 molecular weight was a secretary protein in the system and its expression amounted to 65% of the total protein in the medium supernatant. The TMPFc showed a promotive effect on the growth of Ba/F3-mpl in vitro. A significant portion of the secretary protein existed as dimer, which provided material for studying the dimer in future.
Asunto(s)
Humanos , Secuencia de Aminoácidos , Western Blotting , Fragmentos Fc de Inmunoglobulinas , Genética , Inmunoglobulina G , Genética , Datos de Secuencia Molecular , Pichia , Genética , Plásmidos , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión , Trombopoyetina , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To understand the infectivity and pathogenicity of the plasma of hepatitis E virus (HEV) viremia to primate animals.</p><p><b>METHODS</b>RNA fragment of HEV genotype IV was detected on one healthy donor who was positive for anti-HEV IgM and negative for anti-HEV IgG. Then 10 ml plasma from above donor was transfused to rhesus monkey to observe its infectivity and pathogenicity.</p><p><b>RESULTS</b>Acute hepatitis E was developed in rhesus monkey who accept HEV RNA positive plasma. It was confirmed by virological, immunological, biochemical and histopathological data.</p><p><b>CONCLUSION</b>Acute hepatitis E can be induced by plasma transfusion of HEV viremia, which indicate the possibility of transfusion transmitted hepatitis E</p>