[Expression and preliminary study of HCV F protein gene].
OBJECTIVE
To express HCV F protein gene fragment in E.coli and detect if there is its antibody in HCV patients.


METHODS
RNA of the virus identified as genetype 1b was choosed as template, the F protein gene was amplified by RT-PCR. This gene fragment was inserted into plasmid vector pGEM simple T. F gene was digested by EcoR I and BamH I from pGEM and cloned into plasmid vector pGEX-4T-2. The ligation mixture was transformed into E.coli. TG1 and F fragment expression was induced by IPTG. The expressed protein was purified from lysates with Glutathione Sepharose 4B. The purified protein was used to identify whether there was anti-F antibody in the patients which HCV RNA was positive by ELISA.


RESULTS
After IPTG induction, a positive band about 43 kD was detected. 82 of 120 HCV RNA positive's sera had anti-F antibody by ELISA.


CONCLUSIONS
It is possible to efficiently express the HCV F protein in E.coli. The positive rate of anti-F antibody in HCV RNA positive patients was 68%.
Jiang
Chun-mei
CM
Department of Epidemiology and Biostatistics, Nanjing Medical University, Nanjing 210029, China. jcm80@tom.com
Diao
Zhen-yu
ZY
Cui
Guo-xing
GX
Qian
Wan-hong
WH
Deng
Xiao-zhao
XZ
Xu
Ke
K
Ding
Wei-liang
WL
Tan
Yong-fei
YF
Zhang
Yun
Y
D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D005838:Genotype; D006508:Hepatitis Antibodies / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014758:Viral Core Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
D006508:Hepatitis Antibodies; D014758:Viral Core Proteins; C433031:hepatitis C protein F, Hepatitis C virus