Biology Reference
In-Depth Information
Table 1.
Oligonucleotides Used in this Study
Oligonucleotide
Sequence (5´ to 3´)
HEVBacBg
CGCAGATCTATGGCGGTCGCTCCAGCCC
HEV52Pr
CTGCAGCTATGCTAGCGCAGAGTG
Htg5(0)
CAGCCTGAACTCGCTCCAGAGGA
Htg5(+1)
GCCAGCCTGAACTCGCTCCAGAGGA
Htg3(0)
ATCTTCCGGATCCTCTGGAGCGAG
Htg3(GA)
TCATCTTCCGGATCCTCTGGAGCGAG
Htg3(GG)
CCATCTTCCGGATCCTCTGGAGCGAG
Tag(-52)
CTGCAGCTAATCTTCGGGGTCCTCCGGGGCGAGCT
CAGGCTGTGCTAGCGCAGAGTGG
BglTag
AGATCTATGGCGCAGCCTGAACTCGCTCCAGAGGA
TCCAGAAGATGCGGTCGCTCCAGCCCATGAC
N. (5486) ATGGCG CAGCCTGAACTCGCTCCAGAGGATCCAGAAGATGCG GTCGCT
M A Q P E L A P E D P E D A V A
Site 1. HindIII(5680) GAAGCT CAGCCTGAACTCGCTCCAGAGGATCCGGAAGATGAAGCT TCTAAT
E A Q P E L A P E D P E D E A S N
Site 2. SacII(6245) GGCCGC CAGCCTGAACTCGCTCCAGAGGATCCGGAAGATCGC GGGATA
G R Q P E L A P E D P E D R G I
(5480) N
dORF2
C (7126)
Site 3. Bss HII(6664) ACCGGC CAGCCTGAACTCGCTCCAGAGGATCCGGAAGATGGCGCG CAGGCC
G A Q P E L A P E D P E D G A Q A
Site 4. Sac II(6773) CTCCGC CAGCCTGAACTCGCTCCAGAGGATCCGGAAGATCGC GGTAAG
L R Q P E L A P E D P E D R G K
C .(6973) CTAG CACAGCCTGAGCTCGCCCCGGAGGACCCCGAAG ATTAG
L A Q P E L A P E D P E D *
Fig. 1. Schematic diagram and sequences around the tag epitope insertion
sites in dORF2. Upper rows show nucleotide sequences and lower rows
show the corresponding amino acid sequences. Amino acid numbers relative
to the full-length ORF2 are indicated next to the amino acid before the
inserted amino acids. Nucleotide numbers referring to HEV genome are in
parenthesis. Inserted sequences are underlined. The tag epitope amino acid
sequence is in bold face.
with baculovirus DNA, and the recombinant baculovirus was generated
as described previously. 6 The production and purification of HEV-VLP
was performed as described previously. 6,7
Utilizing any of the insertion sites, the chimeric dORF2 was
expressed at almost equal levels in the cell lysates (Fig. 2A). The anti-
genicity of the tag epitope was maintained in all cases, as shown by
 
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