实施例5:
携带EGFP基因的西门利克森林病毒类病毒颗粒在标记猴神经细胞中的应用,其步骤是:
手术前,无菌的条件下肌肉注射氯胺酮45mg到猕猴(体重4.5kg),然后肌肉注射戊巴比妥135mg和阿托品0.18mg。然后移除面积为7cm×2.8cm的颅骨,剥除硬脑膜以暴露皮层,注射4μl携带EGFP基因的西门利克森林病毒类病毒颗粒(滴度是1.8×108FFU/ml)定位注射到猴脑(猕猴,猴龄12年)到猴脑。当注射器的针头达到目的脑区后,等待2min,然后开始缓慢的注射,每次注射使用时间约为20min,注射完毕后等待约10min,然后再拔出针头。术后3天,每天肌肉注射头孢菌素112.5mg。
采用逆差计时的方式分别在取脑组织前1、2、5、7、10天麻醉猴后,用0.9%(V/V)生理盐水灌流,然后用4%(V/V)多聚甲醛固定,取出脑组织浸泡于4%(V/V)多聚甲醛液中2周,然后将脑组织先置于20%(V/V)蔗糖溶液中1天,然后置于30%(V/V)蔗糖溶液中;将脑组织底部切平,置于底座上包埋冰冻1h后切片;取脑片后使用荧光显微镜观察。在注射携带EGFP基因的西门利克森林病毒类病毒颗粒后2天,可见明显的绿色荧光蛋白的表达,表明携带EGFP基因的西门利克森林病毒类病毒颗粒能够感染猴脑神经细胞,并能在其中复制和表达蛋白(图5),该结果表明携带EGFP基因的西门利克森林病毒类病毒颗粒具有标记非人灵长类动物神经网络的潜力,其标记的速度快速(只需要2天),因此是一种高效的标记非人灵长类动物神经元的工具。
最后,还需要注意的是,上述实施例中的实验方法,如无特殊说明,均为常规方法,并且以上列举的仅是本发明的若干个具体实施例。显然,本发明不限于以上实施例,还可以有许多变形。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形,均应认为是本发明的保护范围。
SEQUENCE LISTING
<110> 中国科学院武汉物理与数学研究所
<120> 一种用于非人灵长类神经细胞快速标记的西门利克森林病毒复制子及应用
<130> 一种用于非人灵长类神经细胞快速标记的西门利克森林病毒复制子及应用
<160> 8
<170> PatentIn version 3.1
<210> 1
<211> 717
<212> DNA
<213> 人工序列
<400> 1
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
cgagctggac 60
ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
tgccacctac 120
ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
ctggcccacc 180
ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
ccacatgaag 240
cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
caccatcttc 300
ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
cgacaccctg 360
gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
cctggggcac 420
aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
gcagaagaac 480
ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
gcagctcgcc 540
gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
cgacaaccac 600
tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
tcacatggtc 660
ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
gtacaag 717
<210> 2
<211> 59
<212> DNA
<213> 人工序列
<400> 2
ccttctcaag ctggcgggag acgtcgagtc caaccctggg ccaatggtga
gcaagggcg 59
<210> 3
<211> 45
<212> DNA
<213> 人工序列
<400> 3
ggatcgacta gtgaactcga gttacttgta cagctcgtcc
atgcc
45
<210> 4
<211> 102
<212> DNA
<213> 人工序列
<400> 4
atgaattaca tccctacgca aacgttttac ggccgccggt ggcgcccgcg
cccggcggcc 60
cgtccctggc cgttgcaggc cactccggtg gctcccgtcg
tc
102
<210> 5
<211> 50
<212> DNA
<213> 人工序列
<400> 5
aatacacaga attctgattg gatccaccat gaattacatc
cctacgcaaa
50
<210> 6
<211> 59
<212> DNA
<213> 人工序列
<400> 6
ctcccgccag cttgagaagg tcaaaattca acagctgccc ggggacgacg
ggagccacc 59
<210> 7
<211> 60
<212> DNA
<213> 人工序列
<400> 7
cagctgttga attttgacct tctcaagctg gcgggagacg tcgagtccaa
ccctgggcca 60
<210> 8
<211> 11471
<212> DNA
<213> 人工序列
<400> 8
gatggcggat gtgtgacata cacgacgcca aaagattttg ttccagctcc
tgccacctcc 60
gctacgcgag agattaacca cccacgatgg ccgccaaagt gcatgttgat
attgaggctg 120
acagcccatt catcaagtct ttgcagaagg catttccgtc gttcgaggtg
gagtcattgc 180
aggtcacacc aaatgaccat gcaaatgcca gagcattttc gcacctggct
accaaattga 240
tcgagcagga gactgacaaa gacacactca tcttggatat cggcagtgcg
ccttccagga 300
gaatgatgtc tacgcacaaa taccactgcg tatgccctat gcgcagcgca
gaagaccccg 360
aaaggctcgt atgctacgca aagaaactgg cagcggcctc cgggaaggtg
ctggatagag 420
agatcgcagg aaaaatcacc gacctgcaga ccgtcatggc tacgccagac
gctgaatctc 480
ctaccttttg cctgcataca gacgtcacgt gtcgtacggc agccgaagtg
gccgtatacc 540
aggacgtgta tgctgtacat gcaccaacat cgctgtacca tcaggcgatg
aaaggtgtca 600
gaacggcgta ttggattggg tttgacacca ccccgtttat gtttgacgcg
ctagcaggcg 660
cgtatccaac ctacgccaca aactgggccg acgagcaggt gttacaggcc
aggaacatag 720
gactgtgtgc agcatccttg actgagggaa gactcggcaa actgtccatt
ctccgcaaga 780
agcaattgaa accttgcgac acagtcatgt tctcggtagg atctacattg
tacactgaga 840
gcagaaagct actgaggagc tggcacttac cctccgtatt ccacctgaaa
ggtaaacaat 900
cctttacctg taggtgcgat accatcgtat catgtgaagg gtacgtagtt
aagaaaatca 960
ctatgtgccc cggcctgtac ggtaaaacgg tagggtacgc cgtgacgtat
cacgcggagg 1020
gattcctagt gtgcaagacc acagacactg tcaaaggaga aagagtctca ttccctgtat
1080
gcacctacgt cccctcaacc atctgtgatc aaatgactgg catactagcg
accgacgtca 1140
caccggagga cgcacagaag ttgttagtgg gattgaatca gaggatagtt
gtgaacggaa 1200
gaacacagcg aaacactaac acgatgaaga actatctgct tccgattgtg
gccgtcgcat 1260
ttagcaagtg ggcgagggaa tacaaggcag accttgatga tgaaaaacct
ctgggtgtcc 1320
gagagaggtc acttacttgc tgctgcttgt gggcatttaa aacgaggaag
atgcacacca 1380
tgtacaagaa accagacacc cagacaatag tgaaggtgcc ttcagagttt
aactcgttcg 1440
tcatcccgag cctatggtct acaggcctcg caatcccagt cagatcacgc
attaagatgc 1500
ttttggccaa gaagaccaag cgagagttaa tacctgttct cgacgcgtcg
tcagccaggg 1560
atgctgaaca agaggagaag gagaggttgg aggccgagct gactagagaa
gccttaccac 1620
ccctcgtccc catcgcgccg gcggagacgg gagtcgtcga cgtcgacgtt
gaagaactag 1680
agtatcacgc aggtgcaggg gtcgtggaaa cacctcgcag cgcgttgaaa
gtcaccgcac 1740
agccgaacga cgtactacta ggaaattacg tagttctgtc cccgcagacc
gtgctcaaga 1800
gctccaagtt ggcccccgtg caccctctag cagagcaggt gaaaataata
acacataacg 1860
ggagggccgg ccgttaccag gtcgacggat atgacggcag ggtcctacta
ccatgtggat 1920
cggccattcc ggtccctgag tttcaggctt tgagcgagag cgccactatg
gtgtacaacg 1980
aaagggagtt cgtcaacagg aaactatacc atattgccgt tcacggaccc
tcgctgaaca 2040
ccgacgagga gaactacgag aaagtcagag ctgaaagaac tgacgccgag
tacgtgttcg 2100
acgtagataa aaaatgctgc gtcaagagag aggaagcgtc gggtttggtg
ttggtgggag 2160
agctaaccaa ccccccgttc catgaattcg cctacgaagg gctgaagatc
aggccgtcgg 2220
caccatataa gactacagta gtaggagtct ttggggttcc gggatcaggc
aagtctgcta 2280
ttattaagag cctcgtgacc aaacacgatc tggtcaccag cggcaagaag
gagaactgcc 2340
aggaaatagt caacgacgtg aagaagcacc gcggactgga catccaggca
aaaacagtgg 2400
actccatcct gctaaacggg tgtcgtcgtg ccgtggacat cctatatgtg
gacgaggctt 2460
tcgcttgcca tcccggtact ctgctagccc taattgctct tgttaaacct
cggagcaaag 2520
tggtgttatg cggagacccc aagcaatgcg gattcttcaa tatgatgcag
cttaaggtga 2580
acttcaacca caacatctgc actgaagtat gtcataaaag tatatccaga
cgttgcacgc 2640
gtccagtcac ggccatcgtg tctacgttgc actacggagg caagatgcgc
acgaccaacc 2700
cgtgcaacaa acccataatc atagacacca caggacagac caagcccaag
ccaggagaca 2760
tcgtgttaac atgcttccga ggctgggtaa agcagctgca gttggactac
cgtggacacg 2820
aagtcatgac agcagcagca tctcagggcc tcacccgcaa aggggtatac
gccgtaaggc 2880
agaaggtgaa tgaaaatccc ttgtatgccc ctgcgtcgga gcacgtgaat
gtactgctga 2940
cgcgcactga ggataggctg gtgtggaaaa cgctggccgg cgatccctgg
attaaggtcc 3000
tatcaaacat tccacagggt aactttacgg ccacattgga agaatggcaa
gaagaacacg 3060
acaaaataat gaaggtgatt gaaggaccgg ctgcgcctgt ggacgcgttc
cagaacaaag 3120
cgaacgtgtg ttgggcgaaa agcctggtgc ctgtcctgga cactgccgga
atcagattga 3180
cagcagagga gtggagcacc ataattacag catttaagga ggacagagct
tactctccag 3240
tggtggcctt gaatgaaatt tgcaccaagt actatggagt tgacctggac
agtggcctgt 3300
tttctgcccc gaaggtgtcc ctgtattacg agaacaacca ctgggataac
agacctggtg 3360
gaaggatgta tggattcaat gccgcaacag ctgccaggct ggaagctaga
cataccttcc 3420
tgaaggggca gtggcatacg ggcaagcagg cagttatcgc agaaagaaaa
atccaaccgc 3480
tttctgtgct ggacaatgta attcctatca accgcaggct gccgcacgcc
ctggtggctg 3540
agtacaagac ggttaaaggc agtagggttg agtggctggt caataaagta
agagggtacc 3600
acgtcctgct ggtgagtgag tacaacctgg ctttgcctcg acgcgacgtc
acttggttgt 3660
caccgctgaa tgtcacaggc gccgataggt gctacgacct aagtttagga
ctgccggctg 3720
acgccggcag gttcgacttg gtctttgtga acattcacac ggaattcaga
atccaccact 3780
accagcagtg tgtcgaccac gccatgaagc tgcagatgct tgggggagat
gcgctacgac 3840
tgctaaaacc cggcggcagc ctcttgatga gagcttacgg atacgccgat
aaaatcagcg 3900
aagccgttgt ttcctcctta agcagaaagt tctcgtctgc aagagtgttg
cgcccggatt 3960
gtgtcaccag caatacagaa gtgttcttgc tgttctccaa ctttgacaac
ggaaagagac 4020
cctctacgct acaccagatg aataccaagc tgagtgccgt gtatgccgga
gaagccatgc 4080
acacggccgg gtgtgcacca tcctacagag ttaagagagc agacatagcc
acgtgcacag 4140
aagcggctgt ggttaacgca gctaacgccc gtggaactgt aggggatggc
gtatgcaggg 4200
ccgtggcgaa gaaatggccg tcagccttta agggagaagc aacaccagtg
ggcacaatta 4260
aaacagtcat gtgcggctcg taccccgtca tccacgctgt agcgcctaat
ttctctgcca 4320
cgactgaagc ggaaggggac cgcgaattgg ccgctgtcta ccgggcagtg
gccgccgaag 4380
taaacagact gtcactgagc agcgtagcca tcccgctgct gtccacagga
gtgttcagcg 4440
gcggaagaga taggctgcag caatccctca accatctatt cacagcaatg
gacgccacgg 4500
acgctgacgt gaccatctac tgcagagaca aaagttggga gaagaaaatc
caggaagcca 4560
tagacatgag gacggctgtg gagttgctca atgatgacgt ggagctgacc
acagacttgg 4620
tgagagtgca cccggacagc agcctggtgg gtcgtaaggg ctacagtacc
actgacgggt 4680
cgctgtactc gtactttgaa ggtacgaaat tcaaccaggc tgctattgat
atggcagaga 4740
tactgacgtt gtggcccaga ctgcaagagg caaacgaaca gatatgccta
tacgcgctgg 4800
gcgaaacaat ggacaacatc agatccaaat gtccggtgaa cgattccgat
tcatcaacac 4860
ctcccaggac agtgccctgc ctgtgccgct acgcaatgac agcagaacgg
atcgcccgcc 4920
ttaggtcaca ccaagttaaa agcatggtgg tttgctcatc ttttcccctc
ccgaaatacc 4980
atgtagatgg ggtgcagaag gtaaagtgcg agaaggttct cctgttcgac
ccgacggtac 5040
cttcagtggt tagtccgcgg aagtatgccg catctacgac ggaccactca
gatcggtcgt 5100
tacgagggtt tgacttggac tggaccaccg actcgtcttc cactgccagc
gataccatgt 5160
cgctacccag tttgcagtcg tgtgacatcg actcgatcta cgagccaatg
gctcccatag 5220
tagtgacggc tgacgtacac cctgaacccg caggcatcgc ggacctggcg
gcagatgtgc 5280
atcctgaacc cgcagaccat gtggacctgg agaacccgat tcctccaccg
cgcccgaaga 5340
gagctgcata ccttgcctcc cgcgcggcgg agcgaccggt gccggcgccg
agaaagccga 5400
cgcctgcccc aaggactgcg tttaggaaca agctgccttt gacgttcggc
gactttgacg 5460
agcacgaggt cgatgcgttg gcctccggga ttactttcgg agacttcgac
gacgtcctgc 5520
gactaggccg cgcgggtgca tatattttct cctcggacac tggcagcgga
catttacaac 5580
aaaaatccgt taggcagcac aatctccagt gcgcacaact ggatgcggtc
gaggaggaga 5640
aaatgtaccc gccaaaattg gatactgaga gggagaagct gttgctgctg
aaaatgcaga 5700
tgcacccatc ggaggctaat aagagtcgat accagtctcg caaagtggag
aacatgaaag 5760
ccacggtggt ggacaggctc acatcggggg ccagattgta cacgggagcg
gacgtaggcc 5820
gcataccaac atacgcggtt cggtaccccc gccccgtgta ctcccctacc
gtgatcgaaa 5880
gattctcaag ccccgatgta gcaatcgcag cgtgcaacga atacctatcc
agaaattacc 5940
caacagtggc gtcgtaccag ataacagatg aatacgacgc atacttggac
atggttgacg 6000
ggtcggatag ttgcttggac agagcgacat tctgcccggc gaagctccgg
tgctacccga 6060
aacatcatgc gtaccaccag ccgactgtac gcagtgccgt cccgtcaccc
tttcagaaca 6120
cactacagaa cgtgctagcg gctgccacca agagaaactg caacgtcacg
caaatgcgag 6180
aactacccac catggactcg gcagtgttca acgtggagtg cttcaagcgc
tatgcctgct 6240
ccggagaata ttgggaagaa tatgctaaac aacctatccg gataaccact
gagaacatca 6300
ctacctatgt gaccaaattg aaaggcccga aagctgctgc cttgttcgct
aagacccaca 6360
acttggttcc gctgcaggag gttcccatgg acagattcac ggtcgacatg
aaacgagatg 6420
tcaaagtcac tccagggacg aaacacacag aggaaagacc caaagtccag
gtaattcaag 6480
cagcggagcc attggcgacc gcttacctgt gcggcatcca cagggaatta
gtaaggagac 6540
taaatgctgt gttacgccct aacgtgcaca cattgtttga tatgtcggcc
gaagactttg 6600
acgcgatcat cgcctctcac ttccacccag gagacccggt tctagagacg
gacattgcat 6660
cattcgacaa aagccaggac gactccttgg ctcttacagg tttaatgatc
ctcgaagatc 6720
taggggtgga tcagtacctg ctggacttga tcgaggcagc ctttggggaa
atatccagct 6780
gtcacctacc aactggcacg cgcttcaagt tcggagctat gatgaaatcg
ggcatgtttc 6840
tgactttgtt tattaacact gttttgaaca tcaccatagc aagcagggta
ctggagcaga 6900
gactcactga ctccgcctgt gcggccttca tcggcgacga caacatcgtt cacggagtga
6960
tctccgacaa gctgatggcg gagaggtgcg cgtcgtgggt caacatggag
gtgaagatca 7020
ttgacgctgt catgggcgaa aaacccccat atttttgtgg gggattcata
gtttttgaca 7080
gcgtcacaca gaccgcctgc cgtgtttcag acccacttaa gcgcctgttc
aagttgggta 7140
agccgctaac agctgaagac aagcaggacg aagacaggcg acgagcactg
agtgacgagg 7200
ttagcaagtg gttccggaca ggcttggggg ccgaactgga ggtggcacta
acatctaggt 7260
atgaggtaga gggctgcaaa agtatcctca tagccatggc caccttggcg
agggacatta 7320
aggcgtttaa gaaattgaga ggacctgtta tacacctcta cggcggtcct
agattggtgc 7380
gttaatacac agaattctga ttggatccac catgaattac atccctacgc
aaacgtttta 7440
cggccgccgg tggcgcccgc gcccggcggc ccgtccctgg ccgttgcagg
ccactccggt 7500
ggctcccgtc gtccccgggc agctgttgaa ttttgacctt ctcaagctgg
cgggagacgt 7560
cgagtccaac cctgggccaa tggtgagcaa gggcgaggag ctgttcaccg
gggtggtgcc 7620
catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
ccggcgaggg 7680
cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
ccggcaagct 7740
gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
gcttcagccg 7800
ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
aaggctacgt 7860
ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
ccgaggtgaa 7920
gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
tcaaggagga 7980
cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
tctatatcat 8040
ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
acatcgagga 8100
cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
acggccccgt 8160
gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
accccaacga 8220
gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
ctctcggcat 8280
ggacgagctg tacaagtaac tcgagttcac tagtcgatcc cgcggccgct
ttcgaaccta 8340
ggcaagcatg cgggcccagt gggtaattaa ttgaattaca tccctacgca
aacgttttac 8400
ggccgccggt ggcgcccgcg cccggcggcc cgtccctggc cgttgcaggc
cactccggtg 8460
gctcccgtcg tccccgactt ccaggcccag cagatgcagc aactcatcag
cgccgtaaat 8520
gcgctgacaa tgagacagaa cgcaattgct cctgctagga gcttaattcg
acgaataatt 8580
ggatttttat tttattttgc aattggtttt taatatttcc aaaaaaaaaa
aaaaaaaaaa 8640
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaactaga
aatcgcgatt 8700
tctagtctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
tattgggcgc 8760
tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
gcgagcggta 8820
tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
cgcaggaaag 8880
aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
gttgctggcg 8940
tttttccata ggctccgccc ccctgacgag catcacaaaa atcgaccctc
aagtcaaagg 9000
gggcgaaacc cgacaggact ataaaaatac caggcgtttc cccctggaag
ctccctcggg 9060
cgctctcctg ttccaaccct gccgcttacc ggatacctgt ccccctttct
cccttcggga 9120
agcggggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
ggtcgttcgc 9180
tccaagctgg gctgtgtgca caaacccccc gttcagcccg accgctgcgc
cttatccggt 9240
aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
agcagccact 9300
ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
gaagtggtgg 9360
cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
gaagccagtt 9420
accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
tggtagcggt 9480
ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
agaagatcct 9540
ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
agggattttg 9600
gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
atgaagtttt 9660
aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
cttaatcagt 9720
gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
actccccgtc 9780
gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
aatgataccg 9840
cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
cggaagggcc 9900
gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
ttgttgccgg 9960
gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
10020
ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
10080
tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
10140
ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
10200
cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
10260
accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
10320
cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
10380
tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
10440
cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
10500
acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
10560
atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
10620
tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
10680
aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
10740
cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
10800
atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
10860
cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
10920
gagcagattg tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattag
10980
gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
11040
caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
11100
acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
11160
ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
11220
gaggatctgg ctagcgatga ccctgctgat tggttcgctg accatttccg ggtgcgggac
11280
ggcgttacca gaaactcaga aggttcgtcc aaccaaaccg actctgacgg cagtttacga
11340
gagagatgat agggtctgct tcagtaagcc agatgctaca caattaggct tgtacatatt
11400
gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
11460
gtgacactat
a
11471