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KMID : 0357419930230020223
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Korean journal of Virology
1993 Volume.23 No. 2 p.223 ~ p.232
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강주현
이영희/윤미리/윤희식/권두한/최용경/박순희/최인성/정노팔
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Abstract
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Human papilloma virus(HPV) type 16 capsid L2 was expressed in E. coli by cloning the L2 open reading frame(1420 nucleotides)into a plasmid pET-3a. The expressed L2 protein was accumulated as inclusion bodies. The inclusion bodies were washed with
the
buffer containing 1M urea and 0.5% Triton X-100 and solubilized in a buffer containing 4M urea and 1mM DTT and then dialyzed in 2M urea. The protein was purified by column chromatographies using Q-Sepharose column followed by Sephacryl S200
column
in a
50mM Tris-Cl buffer, pH 8.0, containing 2M urea and 1mM DTT. The protein was then renatured by applying sequential dialysis protocol and the purity was assessed by 10% SDS-PAGE and immunostaining using polyclonal antibody acquired from TrpE-fused
L2
protein immunized rabbit. The final yield of the purified protein was approximately 13%. The antigenicity of the purified and renatured L2 protein was confirmed by showing the reactivity to rabbit polyclonal antibody against TrpE-fused L2 protein
using
enzyme linked immunosorbent assay(ELISA).
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KEYWORD
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