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. 1998 Apr;4(2):165-168.
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

Cloning and expression of NS3 cDNA fragment of HCV genome of Hebei isolate in E.coli

Fen-Lu Zhu et al. World J Gastroenterol. 1998 Apr.

Abstract

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 M(r) 23800 and an M(r) 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.

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Figures

Figure 1
Figure 1
SDS-PAGE analysis of HCV NS3 fusion and non-fusion proteins expressed in E. coli A: Control HB101/pRX B: Induced HB101/pRXNS3 C: Control HB101/pMY D: Induced HB101/pMYNS3 E: Molecular weight standards
Figure 2
Figure 2
Western blot analysis of HCV NS3 recombinant proteins A: Control HB101/pRX B: Induced HB101/pRXNS3 C: Induced HB101/pMYNS3 D: Control HB101/pMY
See this image and copyright information in PMC

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