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PHRP : Osong Public Health and Research Perspectives

OPEN ACCESS. pISSN: 2210-9099. eISSN: 2233-6052
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Original Article

Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris

Osong Public Health and Research Perspectives 2014;5(5):274-278.
Published online: September 4, 2014

aDivision of Arboviruses, Korea National Institute of Health, Cheongju, Korea

bDivision of Antimicrobial Resistance, Korea National Institute of Health, Cheongju, Korea

∗Corresponding author. wychoi65@nih.go.kr
• Received: August 4, 2014   • Revised: August 18, 2014   • Accepted: August 25, 2014

© 2014 Published by Elsevier B.V. on behalf of Korea Centers for Disease Control and Prevention.

This is an Open Access article distributed under the terms of the CC-BY-NC License (http://creativecommons.org/licenses/by-nc/3.0).

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Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris
Osong Public Health Res Perspect. 2014;5(5):274-278.   Published online October 31, 2014
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Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris
Osong Public Health Res Perspect. 2014;5(5):274-278.   Published online October 31, 2014
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Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris
Image Image Image
Figure 1 Construction of recombinant plasmid pGAPZɑA/93prM-E. (A) Scheme for cloning the 93prM-E fragment into the pGAPZɑA vector. (B) Confirmation of vector and insert by digestion with the restriction enzymes, Bst BI and Xba I. bp = base pairs, E = envelope protein; Lane M = 1 Kb DNA plus ladder; Lane 1 = pGAPZɑA/93prM-E digested with Bst BI and Xba I; prM = premembrane protein; S = the signal peptide of prM.
Figure 2 Western blot analysis on the expression of tick-borne encephalitis virus E protein in Pichia pastoris. Lane 1 = plasmid alone control; Lane 2 = sample from the cell lysate; Lane 3 = sample from the culture supernatant.
Figure 3 Analysis of glycosylation of tick-borne encephalitis virus E proteins in Pichia pastoris transformed with plasmid pGAPZɑA/93prM-E. Samples from (A) the cell lysate and (B) the cell supernatant were treated with Endo H (+) or PNGase F (+) and compared with untreated controls (−) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting.  = E protein;  = deglycosylated E protein; Endo H = endoglycosidase H; PNGase = N-glycosidase F.
Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris