PMID- 28959307
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200930
IS  - 1728-3043 (Print)
IS  - 2322-2921 (Electronic)
IS  - 1728-3043 (Linking)
VI  - 13
IP  - 4
DP  - 2015 Dec
TI  - Analysis of Immumoreactivity of Heterologously Expressed Non-structural Protein 
      4B (NS4B) from Hepatitis C Virus (HCV) Genotype 1a.
PG  - 32-37
LID - 10.15171/ijb.1321 [doi]
AB  - BACKGROUND: Detection of hepatitis C virus specific antibodies is the initial 
      step in chronic HCV diagnosis. HCV NS4B is among the most immunogenic HCV 
      antigens and has been widely used in commercial Enzyme Immunoassays (EIA). 
      Additionally, NS4B, a key protein in the virus replication, can be an alternative 
      target for antiviral therapy. OBJECTIVES: Development of a new method for 
      high-level expression and purification of NS4B coding region was the aim of the 
      report. MATERIALS AND METHODS: Viral RNA was purified from the serum of an HCV 
      positive patient and NS4B coding region was amplified using nested RT-PCR. PCR 
      products were cloned into pET102/D-TOPO expression vector and transformed into E. 
      coli BL21. Induction was performed by adding 1 mM 
      isopropyl-beta-D-thiogalactopyranoside (IPTG) to the culture medium. 
      Immunoreactivity of the purified recombinant proteins was evaluated by 
      immunoblotting and indirect enzymelinked immunosorbent assay (ELISA). RESULTS: 
      The recombinant NS4B protein was expressed and its immunoreactivity was confirmed 
      by ELISA and western blotting. CONCLUSIONS: The directional TOPO cloning provides 
      an efficient and easy platform for heterologous expression of immunoreactive HCV 
      NS4B.
FAU - Savardashtaki, Amir
AU  - Savardashtaki A
AD  - Cellular and Molecular Research Center, Department of Medical Biotechnology, 
      Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
FAU - Sharifi, Zohreh
AU  - Sharifi Z
AD  - Department of Virology, Iranian Blood Transfusion Organization, Tehran, Iran.
FAU - Hamzehlou, Sepideh
AU  - Hamzehlou S
AD  - Cellular and Molecular Research Center, Department of Medical Biotechnology, 
      Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
FAU - Farajollahi, Mohammad M
AU  - Farajollahi MM
AD  - Cellular and Molecular Research Center, Department of Medical Biotechnology, 
      Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
LA  - eng
PT  - Journal Article
PL  - Iran
TA  - Iran J Biotechnol
JT  - Iranian journal of biotechnology
JID - 101276796
PMC - PMC5458682
OTO - NOTNLM
OT  - Hepatitis C Virus
OT  - NS4B
OT  - Recombinant antigen
OT  - Serodiagnosis
EDAT- 2015/12/01 00:00
MHDA- 2015/12/01 00:01
PMCR- 2015/12/01
CRDT- 2017/09/30 06:00
PHST- 2017/09/30 06:00 [entrez]
PHST- 2015/12/01 00:00 [pubmed]
PHST- 2015/12/01 00:01 [medline]
PHST- 2015/12/01 00:00 [pmc-release]
AID - 10.15171/ijb.1321 [doi]
PST - ppublish
SO  - Iran J Biotechnol. 2015 Dec;13(4):32-37. doi: 10.15171/ijb.1321.