TRANSFECTION OF SF9 INSECT CELLS BY RECOMBINANT BACULOVIRUS CONTAINING CORE+1 GENE OF HCV JFH1 GENOTYPE 2A IN ORDER TO EXPRESS CORE+1 PROTEIN

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SAJADIAN FARD FARIDEH SADAT; RAHIMI POONEH

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Journal: JOURNAL OF MICROBIAL WORLD Year:2016 | Volume:9 | Issue:2 (27) Page(s): 85-95

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Title

TRANSFECTION OF SF9 INSECT CELLS BY RECOMBINANT BACULOVIRUS CONTAINING CORE+1 GENE OF HCV JFH1 GENOTYPE 2A IN ORDER TO EXPRESS CORE+1 PROTEIN

Author(s)

SAJADIAN FARD FARIDEH SADAT | RAHIMI POONEH | Issue Writer Certificate

Keywords

HEPATITIS CVIRUSQ1

PROTEIN CORE+1Q1

JFH1 STRAINQ1

INSECT CELLS (SF9)Q1

BACULOVIRUS EXPRESSION SYSTEMQ1

Abstract

Background & Objectives: Recently, a new protein, named Core+1, has been reported to be expressed through a+1 ribosomal frame shift in the Core protein coding region of Hepatitis C virus. The purpose of this study was to design a recombinant Baculovirus vector containing HCV -2a (JFH1) Core+1 sequence, and also, to verify the production of this recombinant Core+1 protein in BACULOVIRUS EXPRESSION SYSTEM.Materials & Methods: Core+1 gene of HCV -2a (JFH1) was synthesized into pUC57 plasmid. The synthesized target gene was sub-cloned into the plasmid pFastBac-HTA. The recombinant vector was used to transform the competent E. coli DH10Bac containing the donor clone. The recombinant Baculovirus bacmid was produced following transposition. Recombinant bacmid was verified by PCR and then was transfected into Sf9 insect cells to package a new recombinant Baculovirus. SDS-PAGE and Western blotting was used to confirm the expression of Core+1 protein in insect cells.Results: Sequence analysis and white-blue colony selection confirmed a successful cloning of the Core+1 sequence of HCV (JFH1) into the pFastBac-HTA vector and transformation of E. coli DH10Bac. Production of recombinant Baculovirus. The HCV -2a (JFH1) Core+1 protein was successfully expressed and confirmed in SDS-PAGE and Western blot.Conclusions: BACULOVIRUS EXPRESSION SYSTEM provides a high yield post-translational modification tool for HCV Core+1 protein expression, which is similar to Core+1 protein produced during natural infection with HCV.

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APA: Copy

SAJADIAN FARD, FARIDEH SADAT, & RAHIMI, POONEH. (2016). TRANSFECTION OF SF9 INSECT CELLS BY RECOMBINANT BACULOVIRUS CONTAINING CORE+1 GENE OF HCV JFH1 GENOTYPE 2A IN ORDER TO EXPRESS CORE+1 PROTEIN. JOURNAL OF MICROBIAL WORLD, 9(2 (27)), 85-95. SID. https://sid.ir/paper/189361/en

Vancouver: Copy

SAJADIAN FARD FARIDEH SADAT, RAHIMI POONEH. TRANSFECTION OF SF9 INSECT CELLS BY RECOMBINANT BACULOVIRUS CONTAINING CORE+1 GENE OF HCV JFH1 GENOTYPE 2A IN ORDER TO EXPRESS CORE+1 PROTEIN. JOURNAL OF MICROBIAL WORLD[Internet]. 2016;9(2 (27)):85-95. Available from: https://sid.ir/paper/189361/en

IEEE: Copy

FARIDEH SADAT SAJADIAN FARD, and POONEH RAHIMI, “TRANSFECTION OF SF9 INSECT CELLS BY RECOMBINANT BACULOVIRUS CONTAINING CORE+1 GENE OF HCV JFH1 GENOTYPE 2A IN ORDER TO EXPRESS CORE+1 PROTEIN,” JOURNAL OF MICROBIAL WORLD, vol. 9, no. 2 (27), pp. 85–95, 2016, [Online]. Available: https://sid.ir/paper/189361/en

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