The construction and primary screening of a phage display library of HCV C and E1 genes evolved with an artificial pattern / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 666-669, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-260638
ABSTRACT
<p><b>OBJECTIVES</b>To construct and screen a primarily phage display library of HCV C and E1 genes evolved with an artificial pattern.</p><p><b>METHODS</b>Two genes of about 1 kb with different genotypes were evolved by DNA shuffling. The re-assembled HCV C and E1 genes were cloned into a phage vector. After being rescued with helper phage M13KO7, a phage display library was constructed. Then the library was screened with anti-C and E1 McAb. Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was carried out on twenty individual phage clones selected randomly to detect their binding and reactive activity with high-titer HCV-positive sera. Normal sera were used as controls.</p><p><b>RESULTS</b>The phage display library of HCV C and E1 genes which evolved with an artificial pattern was constructed. Their capacity amounted to 1.64 x 10(6), and 86 percent of the clones contained C and E1 genes. After four rounds of panning, the phage library was specifically enriched. Twelve positive clones were successfully screened.</p><p><b>CONCLUSION</b>The capacity and diversity of the constructed library are enough for screening. The results demonstrate the superiority of the specific binding and reactive activity and affinity of the 12 phage clones from the HCV positive sera.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
DNA Viral
/
Biblioteca Gênica
/
Proteínas do Envelope Viral
/
Proteínas do Core Viral
/
Hepacivirus
/
Biblioteca de Peptídeos
/
Genética
Tipo de estudo:
Estudo diagnóstico
/
Estudo de rastreamento
Idioma:
Chinês
Revista:
Chinese Journal of Hepatology
Ano de publicação:
2006
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS