<110>    SK bioscience Co., Ltd.

<120>    Method for producing reassortant rotavirus

<130>    OPP20220619KR

<150>    KR10-2021-0070036
<151>    2021-05-31

<160>    18

<170>    KopatentIn 1.71

<210>    1
<211>    1062
<212>    DNA
<213>    Rotavirus strain WC3

<220>
<221>    gene
<222>    (1)..(1062)
<223>    V7 gene segment (AY050272)


<400>    1
ggctttaaaa gagagaattt ccgtctggct agcggttagc tccttttaat gtatggtatt         60

gaatatacca caattctaat cttcttgaca tcgattacat tattaaatta tatcttaaaa        120

tcaataacga gaataatgga ctatataatt tatagatttt tgcttatggt agtgatcttg        180

gccaccataa taaatgcgca aaactatgga gtaaatttgc caattacagg ttcaatggat        240

actgcgtatg caaactctac acaaagtgag ccatttttga catcaaccct ttgtttgtat        300

tatcctgttg aggcatcaaa cgaaatagct gatactgaat ggaaagatac cttatcacaa        360

ctgttcttaa caaaaggatg gccaacagga tcagtgtact ttaaagaata tgctgatata        420

gcggcctttt cagtggaacc acagttatac tgtgattata atttagtttt aatgaaatat        480

gactctacac aaaaactaga tatgtctgaa ttggccgatc ttatattgaa tgaatggctg        540

tgcaatccaa tggacataac gctatattat tatcagcaga ctgatgaggc aaataaatgg        600

atatcgatgg gctcttcttg caacgttaaa gtgtgtccat taaatacaca aacacttggt        660

attggatgtc taataactaa tccagacacg tttgaaacag ttgcgacaac ggagaaatta        720

gtgattacag atgttgtaga tggtgtcaac cacaaattaa acgtcacaac ggcaacgtgc        780

accatacgca actgtaaaaa gttaggacca agggagaacg tagcagtcat acaggtaggc        840

ggcgcgaaca ttttagacat cacagctgat ccaacaacta caccacagac agagagaatg        900

atgcgaataa attggaaaaa atggtggcaa tgtttttaca cggtagtgga ttacgtcaat        960

caaataattc agacagtatc caaaagatct agatcgctta attcgtcggc gttctactat       1020

agagtgtagg tgcatcttag attagaattg tatgatgtga cc                          1062


<210>    2
<211>    1062
<212>    DNA
<213>    Rotavirus strain Brb9

<220>
<221>    gene
<222>    (1)..(1062)
<223>    VP7 gene segment (GU565090)


<400>    2
ggctttaaaa gagagaattt ccgtttggct agcggatagc tccttttaat gtatggtatt         60

gaatatacca cagttctatt ttatttgata tcgttcgttc ttgtgagtta tattctgaaa        120

actatgataa agataatgga ctatttaatt tatagaataa catttgtaat tgtagtatta        180

tcagtattat cgaatgcaca aaattatgga ataaatttgc caattactgg atctatggat        240

acagcatatg ctaactcaac acaagacaat aattttttat cttcaacttt atgtctatat        300

tatccatcag aagctccaac tcaaattagt gacactgaat ggaaagatac actatctcaa        360

ctgtttttaa ccaaaggatg gccgacaggt tcagtttatt ttaatgaata ttcaaacgtt        420

ttagaatttt ccatcgaccc aaagctatac tgtgattata atgttgtgtt aattagattc        480

gcttctggtg aggagttgga cgtatctgaa ttagctgatc taatactcaa tgagtggtta        540

tgtaatccaa tggatataac attatattat tatcaacaaa ctggagaggc aaataagtgg        600

atatcaatgg gatcatcatg taccgttaaa gtgtgtccat taaatactca gacattagga        660

attggatgtc aaacgacaga tactgctact tttgaaacag ttgctgacag cgaaaaattg        720

gcattaattg atgttgttga caacgtaaat cataaattag atgttacatc tactacgtgt        780

acaatacgga attgtaataa actaggacct agagaaaatg tggctataat acaggttggc        840

ggttctaata tattagatat aacagctgat cccacaactt ctccacaaac agaacgaatg        900

atgcgcgtaa actggaaaaa atggtggcaa gtattctata ctgtagttga ttacattaat        960

caaatagtaa aagtaatgtc caaaagatca agatcgttag attcgtcatc tttctattat       1020

agagtataga tatatcctaa aatagaactg tttgatgtga cc                          1062


<210>    3
<211>    21
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    miRNA target site 1


<400>    3
caattctaat cttcttgaca t                                                   21


<210>    4
<211>    21
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    miRNA target site 2


<400>    4
ttgtttgtat tatcctgttg a                                                   21


<210>    5
<211>    21
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    miRNA target site 3


<400>    5
tggtgtcaac cacaaattaa a                                                   21


<210>    6
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA top strand against target site 1


<400>    6
tgctgatgtc aagaagatta gaattggttt tggccactga ctgaccaatt ctacttcttg         60

acat                                                                      64


<210>    7
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA bottom strand against target site 1


<400>    7
cctgatgtca agaagtagaa ttggtcagtc agtggccaaa accaattcta atcttcttga         60

catc                                                                      64


<210>    8
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA top strand against target site 2


<400>    8
tgctgtcaac aggataatac aaacaagttt tggccactga ctgacttgtt tgttatcctg         60

ttga                                                                      64


<210>    9
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA bottom strand against target site 2


<400>    9
cctgtcaaca ggataacaaa caagtcagtc agtggccaaa acttgtttgt attatcctgt         60

tgac                                                                      64


<210>    10
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA top strand against target site 3


<400>    10
tgctgtttaa tttgtggttg acaccagttt tggccactga ctgactggtg tcacacaaat         60

taaa                                                                      64


<210>    11
<211>    64
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    pre-miRNA bottom strand against target site 3


<400>    11
cctgtttaat ttgtgtgaca ccagtcagtc agtggccaaa actggtgtca accacaaatt         60

aaac                                                                      64


<210>    12
<211>    1228
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    DNA template for BrB9 VP7 mRNA in vitro transcription


<400>    12
gaattctaat acgactcact actataggct ttaaaagaga gaatttccgt ttggctagcg         60

gatagctcct tttaatgtat ggtattgaat ataccacagt tctattttat ttgatatcgt        120

tcgttcttgt gagttatatt ctgaaaacta tgataaagat aatggactat ttaatttata        180

gaataacatt tgtaattgta gtattatcag tattatcgaa tgcacaaaat tatggaataa        240

atttgccaat tactggatct atggatacag catatgctaa ctcaacacaa gacaataatt        300

ttttatcttc aactttatgt ctatattatc catcagaagc tccaactcaa attagtgaca        360

ctgaatggaa agatacacta tctcaactgt ttttaaccaa aggatggccg acaggttcag        420

tttattttaa tgaatattca aacgttttag aattttccat cgacccaaag ctatactgtg        480

attataatgt tgtgttaatt agattcgctt ctggtgagga gttggacgta tctgaattag        540

ctgatctaat actcaatgag tggttatgta atccaatgga tataacatta tattattatc        600

aacaaactgg agaggcaaat aagtggatat caatgggatc atcatgtacc gttaaagtgt        660

gtccattaaa tactcagaca ttaggaattg gatgtcaaac gacagatact gctacttttg        720

aaacagttgc tgacagcgaa aaattggcat taattgatgt tgttgacaac gtaaatcata        780

aattagatgt tacatctact acgtgtacaa tacggaattg taataaacta ggacctagag        840

aaaatgtggc tataatacag gttggcggtt ctaatatatt agatataaca gctgatccca        900

caacttctcc acaaacagaa cgaatgatgc gcgtaaactg gaaaaaatgg tggcaagtat        960

tctatactgt agttgattac attaatcaaa tagtaaaagt aatgtccaaa agatcaagat       1020

cgttagattc gtcatctttc tattatagag tatagatata tcctaaaata gaactgtttg       1080

atgtgaccgg gtcggcatgg tcccagcctc ctcgctggcg ccggctgggc aacattccga       1140

ggggaccgtc ccctcggtaa tggcgaatgg gacgcatagc ataacccctt ggggcctcta       1200

aacgggtctt gaggggtttt ttggatcc                                          1228


<210>    13
<211>    25
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    WC3V7_F primer


<400>    13
tcttgacatc gattacatta ttaaa                                               25


<210>    14
<211>    29
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    WC3V7_R primer


<400>    14
catatctagt ttttgtgtag agtcatatt                                           29


<210>    15
<211>    24
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Brb9V7_F primer


<400>    15
ttcgttcttg tgagttatat tctg                                                24


<210>    16
<211>    20
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    Brb9V7_R primer


<400>    16
gtccaactcc tcaccagaag                                                     20


<210>    17
<211>    22
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    WC3V4_F primer


<400>    17
gtcgttcgat ctaccagtag ga                                                  22


<210>    18
<211>    25
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    WC3V4_R primer


<400>    18
gtcactatat ggccttacct caact                                               25
