Zhu FL, Lu HY, Li Z, Qi ZT. Cloning and expression of NS3 cDNA fragment of HCV genome of Hebei isolate in E. coli. World J Gastroenterol 1998; 4(2): 165-168
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Dr. Fen-Lu Zhu, Department of Microbiology, Second Military Medical University, Shanghai 200433, China
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World J Gastroenterol. Apr 15, 1998; 4(2): 165-168 Published online Apr 15, 1998. doi: 10.3748/wjg.v4.i2.165
Cloning and expression of NS3 cDNA fragment of HCV genome of Hebei isolate in E. coli
Fen-Lu Zhu, Hao-Ying Lu, Zhuo Li, Zhong-Tian Qi
Fen-Lu Zhu, Hao-Ying Lu, Zhuo Li, Zhong-Tian Qi, Department of Microbiology, Second Military Medical University, Shanghai 200433, China
Fen-Lu Zhu, male, born on 1963-11-01 in Tang County, Hebei Province, Han nationality, graduated from the Academy of Military Medical Sciences and was offered the Ph.D. degree in 1994, having 8 papers published.
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Fen-Lu Zhu, Department of Microbiology, Second Military Medical University, Shanghai 200433, China
Telephone: 86·21·65347018 ext 71349
Received: September 21, 1997 Revised: February 22, 1998 Accepted: March 28, 1998 Published online: April 15, 1998
AIM: To obtain greater antigenicity of HCV NS3 protein.
METHODS: The HCV NS3 cDNA fragment was amplified by reverse transcription polymerase chain reaction from the sera of the HCV infected patients. The DNA sequence was determined by dideoxy-mediated chain termination method using T7 polymerase. HCV NS3 protein was expressed in E. coli.
RESULTS: Sequence analysis indicated that the HCV isolate of this study belongs to HCV-II; SDS-PAGE demonstrated an Mr 23800 and an Mr 22000 recombinant protein band which amount to 14% and 11% of the total bacterial proteins separately. Western blotting and ELISA showed NS3 protein possessed greater antigenicity.
CONCLUSION: Recombinant HCV NS3 protein was expressed successfully, which provided the basis for developing HCV diagnostic reagents.