[The construction and primary screening of a phage display library of HCV C and E1 genes evolved with an artificial pattern].
OBJECTIVE
To construct and screen a primarily phage display library of HCV C and E1 genes evolved with an artificial pattern.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Zhao
Fu-tao
FT
PLA Center of Diagnosis and Treatment for Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
Jia
Zhan-sheng
ZS
Li
Jin-ge
JG
Wang
Chun-yu
CY
Wei
Xin
X
Li
Guang-yu
GY
Bai
Xue-fan
XF
D004279:DNA, Viral / Q000235:genetics*; D015723:Gene Library; D016174:Hepacivirus / Q000235:genetics*; D019151:Peptide Library; D014758:Viral Core Proteins / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics*
D004279:DNA, Viral; C087035:E1 protein, Hepatitis C virus; D019151:Peptide Library; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins