CN113684281B - 一种采用snp分子标记技术鉴别润州凤头白鸭黑色喙性状的方法 - Google Patents

Abstract

本发明公开了一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙性状的方法，所述分子标记位于润州凤头白鸭基因组的11号染色体上，核苷酸序列如SEQ ID NO.1所示；所述分子标记位于第28397位点，突变碱基为A或T，同时利用该分子标记设计引物（SEQ ID NO.2和SEQ ID NO.3）进行PCR扩增检测。将所得扩增产物进行直接测序后判定受测个体基因型，可以确定润州凤头白鸭黑色喙这一性状，为有目的地对凤头白鸭喙色性状进行选择提供依据，提高润州凤头白鸭黑色喙的一致性。规范我国鸭遗传资源的生产经营，可以为润州凤头白鸭遗传资源保护提供有效保障，支持和引导鸭遗传资源监测评估。

Description

一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙性状的 方法

技术领域

本发明涉及一种鸭喙色的鉴定方法，具体是一种采用SNP分子标记技术鉴别润州凤头白鸭喙色的方法，属于畜牧学、分子生物学领域技术领域。

背景技术

对于畜禽而言，喙色性状是重要的品种特征之一。对于镇江润州地区发现的遗传资源群体--润州凤头白鸭而言，黑色喙则是其重要的外观特征。然而在长期利用传统的育种方法对该资源群体进行抢救性收集与整理中发现，经过多代常规的表型选育，后代喙色仍会发生性状分离，润州凤头白鸭群体的喙色表型始终未达到完全统一，严重影响群体的一致性。准确、高效的喙色选育方法，是润州凤头白鸭遗传资源保护亟待解决的关键问题之一。

SNP(Single Nucleotide Polymorphism,SNP)，称为单核苷酸多态性，是第三代遗传标记，普遍地存在于所有物种基因组染色体上。研究发现，位于基因内部的SNP有可能直接影响蛋白质结构或表达水平，最终导致表型的差异，可作为重要分子标记，用于遗传资源的保护和重要经济性状的选育。

发明内容

本发明的目的是提供一种鉴定润州凤头鸭喙色的特异性分子标记，具体是一种采用SNP分子标记技术精确区分润州凤头白鸭喙色的方法，利用分子标记对鸭遗传资源进行特异性鉴定。

本发明的目的通过以下技术方案实现：一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙性状的方法，其特征是，所述分子标记位于润州凤头鸭基因组的11号染色体上，核苷酸序列如SEQ ID NO.1所示；所述分子标记位于第28397位点，突变碱基为A或T；鉴别时，包括以下步骤：

步骤1)、提取鸭的基因组DNA；

步骤2)、针对MITF-28397设计特异性扩增引物SEQ ID NO.2和SEQ ID NO.3，再以步骤1)的DNA为模板进行PCR扩增反应，获得PCR产物；

步骤3)、将步骤2)得到的PCR产物进行基因序列测序，通过基因测序序列和峰图判定鸭羽色基因型。

进一步的，所述步骤1)中待测鸭个体生物检材的基因组DNA为鸭翅下静脉血样基因组DNA。

进一步的，所述步骤2)中特异性扩增引物序列为：

MI-F1：TCCCCATTCTCCAAAGGTTAGC

MI-R1：TCCCCGTTGTCCCCACTTATTA

进一步的，统计步骤3)中获得的测序结果，判定鸭喙色的对应基因型：

若基因型为AA，表示受测个体为黑色喙纯合野生型基因型；

若基因型为AT，表示受测个体为黑色喙杂合基因型；

若基因型为TT，表示受测个体为黄色喙纯合基因型。

有益效果：本发明通过使用分子标记技术可以实现对润州凤头白鸭喙色性状的快速鉴定，操作简便且结果可靠，为润州凤头白鸭的遗传资源保护提供理论参考。同时为有目的的对润州凤头白鸭喙色性状进行选择提供科学依据，仅需一个世代的检测即可完成育种群在目标喙色性状(黑色)上的纯化。

附图说明

图1为鸭黑色喙和黄色喙个体基因型鉴定图。

具体实施方式

下面结合实施例、附图以及附图说明对本发明做进一步详细说明，所述是对本发明的解释而不是限定。

一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙性状的方法，具体步骤如下：

1.材料与方法；

1.1实验材料；

选择润州凤头白鸭黑色喙个体23只，黄色喙个体21只，其他鸭品种黑色喙个体59只，黄色喙个体70只。

1.2受测个体基因组DNA提取；

每只鸭个体于翅静脉处采血，用传统酚-氯仿法提取基因组总DNA，测DNA浓度和纯度，将DNA母液分装稀释至50ng/μl，于-20℃冰箱保存。

1.3以所提取基因组DNA为模板，通过PCR引物进行常规PCR反应；

1.3.1扩增引物

MI-F1：TCCCCATTCTCCAAAGGTTAGC，

MI-R1：TCCCCGTTGTCCCCACTTATTA；

1.3.2PCR反应体系及反应条件；

a.PCR具体反应体系见下表：

PCR反应体系

b.PCR具体反应条件见下表：

PCR反应条件

1.4将得到的PCR扩增产物送至擎科生物股份有限公司进行测序；

1.5使用ContigExpress软件针对测序结果进行统计分析，获得各群体基因型频率分布。

2.结果

根据图1判定待测个体在该位点的基因型。纯合黑色喙个体基因型为AA；杂合黑色喙个体基因型为AT；纯合黄色喙个体基因型为TT。

针对MITF-28397位点在不同喙色鸭个体的基因型进行统计分析(表1)。其中，在润州凤头白鸭黑色喙群体中发现52.17％纯合野生型基因型个体和47.83％杂合基因型个体(n＝23)，而在黄色喙群体中无纯合野生型个体，突变基因型和杂合基因型个体分别占61.90％和38.10％(n＝21)。在其他鸭品种中，黑色喙群体发现69.49％的个体为纯合野生型，杂合基因型个体占30.51％(n＝59)；而在黄色喙鸭群体发现100％突变基因型(n＝70)。

表1不同喙色类型(黑色、黄色)个体基因型

序列表

<110> 扬州大学

<120> 一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙性状的方法

<160> 3

<170> SIPOSequenceListing 1.0

<210> 1

<211> 39457

<212> DNA

<213> 鸭(mallard)

<400> 1

ctaacaagca tggtcggtgt cctccatgct cacactgctc cgcctgctac ttgttttcga 60

tgctccggga gacacggagg aaagtagcgg gtcagttact cctacgggag agagggtatc 120

gtccatcaag atatcttcca gtttggatcc catttttgta ggaacactgt aagtgccggt 180

cggttcagtc atgtttccga ggttgtcact gaaggtgatg gtaccgtcag tgagatcaag 240

ggtggtcgtg caagacaggt ctgtgtggtg cggcatgatg tcttggttgc agttgtccag 300

cgcgggttct tgtttgatga ccctgttgac catatcaggg gagcaaaggc cggtggatgg 360

cacaagggac agtccatgtg cccgagcttg catctcaagt tcctgtgaag gaggataaag 420

gagaaattaa aaataataaa aaaaaaatgg aagcaacatt tgtcatctta caagcttgtg 480

tattatgact tacagttctc ttctgttaga taggaaatgt ttcctccctt tccaatggtg 540

acaaacattt gcaccctgta cacttctacg ctgcagattt tcaaccaaat aacattctct 600

tgaagaagtg gataaatgct accaaacaca ggtccttcac agttctcaga acctacccag 660

acacgaaacc cagtgccctg cagaacagac atgttgtccc tcagactcct ttctcaaacc 720

agtgccaggg tagggaagct tggcgtcatg ttggggaagg aaagcatttc aaaagttgac 780

gaaaatgaaa cagcagaagt gaacacttag tgcttttcag aagttacgta tgaaagcaca 840

gtgtgtctat acaaggtagc tacttctttc tgagacactg caagggacat tgcacaccct 900

acagctctgg ttaggaggaa agtgaagata tcagcccaat ctgagcaaca ctactcctgc 960

aataaaaaca tacaggaagg ttacaggaag actttagaat catagaatca tagaatatcc 1020

tgagttggaa gggaccctta aggatcatca agtccaactc ttgacaccgc acaggtctac 1080

ccaaaagttc agaccatgtg actaagcgca cagtccaatc tcttcttaaa ttcagtcagg 1140

ctcggtgcag tgaccacttc cctggggagc ctgttccagt gtgcaaccac tctctctgtg 1200

aagaacttcc tcctgatgtc aagcctaaac ttcccctgcc tcagcttaac cccgttcccg 1260

cgggtcctgt cgctagtgtt aatagagaaa aggtctcctg cctctcgaca cccccttacg 1320

aggaagttgt agactgcgat gaggtctccc ctcagcctcc tcttctccag gctgaacagg 1380

cccagtgccc tcagccgttc ctcgtacgtc ttcccctcca ggcctttcac catcttcgta 1440

gccctcctct ggacactctc caacagtttc atgtcctttt tatactgtgg tgcccagaac 1500

tgcacacagt actcgaggtg aggtcgcacc agcgcagagt agagcgggac aatcacctcc 1560

ctcgacctgc tagcgatgcc gtgcttgatg caccccaggg tatggttggc cctcctggct 1620

gccagggcac actgctggct catattgaac ttgctgtcta ccacgaccct cttctaggct 1680

gctctccagt gtctcatcgc ccagtctgta cgtgcagcca gggtttcccc gtcccaggtg 1740

caggacccgg cacttgctct tattgaactt catgcggttg gcgatcgccc agctctccaa 1800

cctatccaga tccctctgca aggcctttcc accctcattc gagtccacaa ctcctccaag 1860

tttggtgtca tcagcaaact tgctcaaaat accttctatt cctacatcca gatcgtttat 1920

aaaaatattg aaaagtaccg gccctaaaat ggagccttga gggaccccac tagtgaccgc 1980

ccgccagcct gacgcagcgc cattcaccat aaccctttgg gccctgcccg ttagccaatt 2040

gctcacccat catatgatgt ttttatttag ctgtatggtg gacattttgt ccagtaggat 2100

cctatgggag accgtgtcaa aagccttgct gatgtccaaa aaaatcacat cagctggttt 2160

cccttggtcc accatacggg tgatcttatc ataaaaggaa atcaggttag ttaggcagga 2220

cctgcccttc acaaacccat gctggctggg accaatgact gctttgtccc ccaggtgcgc 2280

ctcaataagt tcgagaacca tcttctccat gattttacca ggcactgacg tgagactgac 2340

aggcctcact tcacatgcaa cttcacatgc aactagactt cacatgcaac taggattctt 2400

tgaatcttta ggaaaggctt aaatcccacc ttcacgttta atcgcaggca catttgactg 2460

aaggaagata ataaaagaat gcacttaaat gtcctgtttc aaatacagct gcaagacagg 2520

ttttttgctt ctacttatgc attatattaa gcaatcacaa tcactgttgt gattgcactg 2580

ttcctggata ttcttatcac ggaggttgat taacagcctc ttccatttcc aggctaagac 2640

aacaagcttg caatggcaca ctgcaacagc cacagagaag gtggaaaaga agcacttcca 2700

gcagctaatg tgccaaaatc agaatacaga ggaattcctt catcagaacc aacataaaaa 2760

gggaaaatac agagctcagg ttatttactg gcagatgtat tacccattat tccagggttg 2820

tttgtgtttt tttctcattg aaaagggagc cggagcagat gggtaatgca aaggaagatg 2880

aaagatgctt aaaaagcaat ctgacttcac tttgaaatac cactgcttga tgttgttctg 2940

cagaagggcc atgcttcttt atcttctcac ttgagcaagc agattgctca gcgaggtaca 3000

aaagctacgc tgaagctcat gccagcctgt tctattttaa atctgtttgc tgccagactt 3060

gactctcagg tatttcaagc ttcgttatct actctgtgga ttaactggtt cagggctctc 3120

ctgtatccag ccctgcgcac agcaacctgc agaagggctc agctgatcgc gctgcaagtg 3180

agcactaaca caagagtatc tgtggctcac agtccagcag catcgctgtc tttgtgttat 3240

gtgttcttac acccagttct acacatttct tgaattaacc ttttttgcag agtggtgtat 3300

atccagctat cctggaaaac agggttgata acaaaagctg tacaggcttc taggataagg 3360

ttacctatcc ggtttgaggt ggaggcaagg gattttatga gagaaatgaa gttcccaccc 3420

caaaactgct gtgtaataca gcctactggg aatctgggtg aagctaagaa tgcaacagcc 3480

catgatgaag acagaagaag gagggggaag atatattcat gccccccact gctccctcca 3540

gtgttgagga ggaaagctgt tctaatccta cagtctttaa ggaacagaga agaacaaatt 3600

acagctcctt gaacagaaag catcaacaac cacaaaaaca gccctacatc catagttcct 3660

ggttgtcctg cttcaaaaaa ctcttggggc tctcacattc aggctgcaac tcctctcaga 3720

ctcaaagcag actcaacttt taacacagcc gccaggaacc caatgctgca gtgacaacta 3780

tacttcccgc tgcccacttt gacctgtgag caatgtttac cctgcgaaca aaaaagcaga 3840

tgacagactc ctgcagaatg actcctgctg tcagagaaag gctcagatgt ggaagactcc 3900

agactgcgtt gcctctctga caggcagggc ctacattgtc ctgacagaat taaggaaacc 3960

cacaagcaaa acagattttg ttcattgcta ctttcagaaa cagcttcaga agaggacaca 4020

ccactacagc tccaagtcca caaaactccg caggcacaac taaagcacaa cagacagctg 4080

aactgaactt gattactatc aacaaataga tcatacgtga agtcctggtt ctggtctggc 4140

actcagaaaa aaatattact gtatagatct tttttatagt atggcttgca acttatgtta 4200

gctatcctag aacacaacca gaccagccac aagtgcttta acagaatact gaggagttta 4260

ggggccaggg aggagtgggc agaggagggc atctggttat tttttggtgt gattttattt 4320

attttaacta atgtatcctg ataattctgg aaaaattctg cagacatctc agtacagggt 4380

cactattaat taaaacagtg gctccaccta acccaaatcc ttataaagga ccttcagttg 4440

catctgtttg cagccacaca ctacaacatc aagagcactt gcaggttaag tatccacaaa 4500

cctgtattct gagcagcaga tgcctgttgg catgttccaa tttcttctgc ctgttctcaa 4560

gttcctttgt gcgttgctgt tctctttgaa gcttacgaat gtagtccact gatgctttca 4620

gaatagttcc cttattccag cgcatatccc ttcgaaatca acacaaaagg agaggttaag 4680

gctcccgtgg ggacttcacg ctgtagccag taccttcaga gcagcagctg tgattccacc 4740

tcacgctatg taacagccag ctaatacaat attcctccca agctgatgaa tgcagcaggc 4800

ttttgaaatg cactagaggg ggccagagca ctgctattgc aaggcagcga gaatcgtccc 4860

agctgaagga gagagcccag gcgctgctca gtggtttggg gctccccaca gccctctggg 4920

ccagagggat gagccccggc cttctgcaag cactgcaaag actgaagacc aagggctcct 4980

ttcattgcct tctgtgaggc tgcacaaaaa ccctacagga caccactcaa accatattac 5040

ttgctgtctg ggcgatgcct gatttcaggc agggtaatta aacgactgca ggggaactcg 5100

aattcataaa gaaccttata ggaattggtg tttctgtggg aggaagaccc gaggccgtgc 5160

agctcccaag cagcagcaag caacagtcct cctggtggca ctgtgccaca gcggggtccc 5220

agcaggaaga gcagagccca gcacagtgtg tttggatgct ttacgcttaa ctcttgctaa 5280

aaatataaat gctgtgttgc aaccgaggtc tgaccttcag tgaggggggt acagggctcc 5340

catattgccc ctcccacatt attcctacac cacacaaaag ggtgcctgca ttgtcttctc 5400

ccctgtgctg actttttttt ttatataagg agaaagttta gcacagagtg tacagaccaa 5460

agatccacct gttctgtagg ctgatgatgt gcaacattac cttgccttaa aggaagctaa 5520

agagcctctc taaatagtat ttattagtta aaataagaaa aagtctgccc caaagcctca 5580

ctgggcactg attcacattt aagtgacatt ttcaagctcc agagcaagag aggggttcat 5640

tgctatcgaa ccagcattag gtaaatgcca agacaaattg ataacactga aattacctaa 5700

aaagaactac aacccgcctt tctgggaaat taccgacaaa cacagcaaat tatcagttta 5760

acagacacag acctacagtc tttttttctt tcatatctct cttggaggga tggagaatgt 5820

gacggcttcc cagaggaact gccctccact tctacaaaaa gaaggatctg ttctgtcttg 5880

ccaagctatg acatctaata aattcaaagg atcgcttagc aagacaagca gctatgtagt 5940

cagctgagca gctggactag actatgcgtt cacatcctga cagctctctt gctggtacgg 6000

cggtcactca cagcatcagc cccaacgctg caaaagcaca gggccctacc aagatgctga 6060

gctgctgcca gggttctgtg actattttgt gacaaaagta atcaaattaa gaaccttcaa 6120

acacttctac tgccacagaa agccaacatc aaaagagaaa attcacgatt ttatgatttt 6180

atctccccta agatttcttt taaaaacaac caccaccacc ctcccgtcca cagactcctg 6240

ggcacatcat acttacgggt catttgactt gggtatcaaa gtgcctagtt cttttatacg 6300

atcgttaata ttgaatcttc ttcttcgttc aactttagga aaaacagcac agatgttaca 6360

agtcatatac acttaattac aaagtataaa gcttttaatt aaagatttct ttaagtgctt 6420

ttgaaattgt tgcctgaagc ttaaaatcta gtgacaatat agtaatttat tttgctcttc 6480

tacacataga gacagaagtt gtgtttacgc tttaaaaatg tgggactgtt gacgtggcaa 6540

gatctaaagt gagactctaa aaaatacgat acggctgtgt tagcacaaga ctccaataaa 6600

aataacgaaa agcgtttgtc ttccaagggg aaagtatcac aaaaatttgg agacaaaagt 6660

ttattgctca ggaaaatccc aaagcaggaa aagggacaaa aagtataggc atgtttatgt 6720

attctttaag attcaacact taagagtaca taaaaacaat caaacctaaa gatcaacttt 6780

cttgcattag cttttcatgt tattacaaaa gcttaccaaa aaggcctctg aatagcctaa 6840

ggcataaaag agcacctttt tttctattaa tacaaaagac tcttaagttg tcttttgata 6900

cttaaatcat aagatgctca ttgttctatt ttccttttct aaactcagaa atgagtgctc 6960

ccttcaaact ggaatttata aaattgtcac agtgaggatg tgcactggtg tttaaagtca 7020

taagctaact gcctacctta aattgtagtg gttttgttat gttttgtttt taaaaagcaa 7080

aaaatcacaa ggtgaatttg ttgaaagcaa ttggatggga tgttcccagt aattttactg 7140

gttaacacca cgtctgagcg catacttcca agaagtccag ataaacagat cgttacttgg 7200

aattagacca cccagaccat ttcacttttt tggaaaacgg tttaaaaatt gaagagtaga 7260

tttaagtaca cacttcagga gtatcagggg catttgatct atgggcataa cataatgaac 7320

cagtattaga cggagtggct tattcagatt taaaatgctc acattccaac acttaagaac 7380

aactatgatc attattaaaa ttggtgtaaa tagcaaagaa ccaaacttac tcaagttgtg 7440

attgtctttc ttttgcctct ccttagccaa tgctcttgct tctgactctg taggaaaaat 7500

acatgctgtg catgtgaatg aggtaattag aattaaagta attcaaacaa catttcagtt 7560

tgtgaaactt ttgtgaagga agcctgatca gcaactggca aacaaatcat tacagccttt 7620

ccgtaaagtg agaagggcct gtcacataca ttgtgtttac tgaaatccag agctggtatt 7680

caactttacc cctgcacctg cagagctttt gttcccacac ttcctatgct gacttctgtg 7740

atgagaacgt ttcccacgga ggccctgatt cagcaaggta tttgaggaca acaaattgct 7800

gaaatccaac atacctgctc ctacgtacac atgcagacaa catgcttatg taccacacag 7860

aggctgaacc agaggaacaa catgcttact gcattgctac ctgcacagga tgtgaagcag 7920

ccaggagtca gcagctccgc ttgctggaat tgagactatg tccttttgtg gcaacatgtg 7980

cacatcacca cagcactcaa ggtacggact taagtgacct actaaaaatg ctgtgttttc 8040

cagctatatt aattgttccc attatatcaa aaaagttttt aaaaaatcaa cacctttatt 8100

ggcaaatctc agatcataac tgttcttaac tttggtccac acatttatta ctcagacttg 8160

agcacgtata gtcaggcaaa cacataacta agcttgttat ttttcggttc tcatttgatc 8220

ttccatttcc caaactattt caactacttg tttgttctat ccattttaat ttctgaaaaa 8280

aaataaatga ggtgaaatcc ttaacccttt ttccctgttg cacttaaatt acaaaagcag 8340

caaacattag ttttgcagta gatcagtgag gacaagtgaa cctgtgatct attatcacac 8400

attcctttat gggaagaaaa cccttcactc aaacaatttt attttctgta ttattttctt 8460

tttatgcaat atggattatg tcccattcaa gccacagtaa ctggtgacga gggtttattt 8520

cccttggagt caccgtttgt gatagtcaaa ggcttggcca gatcaatgtc aaagtgactg 8580

ctgttaaatt tcagtttaat tcctaaactt aggacaagtt aaatgcaaaa cactttttct 8640

cctaatctat ggtagcagat aactaaattg cttgtttaca catgtgaagg tagcacatag 8700

atctgaacta agctaaatgt ctattttgac acactgagga cttctgtaag accacttcat 8760

tcactaattt cccctctttc tgccttcttt gcccaatgtc ttacacaatc acctaaagtg 8820

aatccaccag cctagacaca tggcaagttg agtgtttagc ttgcagtcta cagccatttg 8880

tagctttctg taactgaggc aatgaagctg tctgtaatgt aggaacctga gtagatctca 8940

tcacctccca ccctcctctt aacatagttc tacatgctgt tgaagttttt ctttctctag 9000

cagccactag tcattttgca ctttgtgtat cttcctttgg cgcagagctc aaattctgcg 9060

cactcataca aaatgcaaat atccaccagc tcagagacgt ggcatttcct ccagagaagc 9120

tcactcctgg gctattaaaa ctgctctact taaactaatt cctgtccaga acctgctttg 9180

ccactgagtc agcctgtgag ttttcacaac agaaagaaga gagacaagaa ataaactaca 9240

cctctaccag gtcagcacac agcatgactc tcagttgggc ttaagagcaa taatttctgc 9300

gtattaatat gattttgtca ctggttctag aatggatctc catttgttca tttgcagaca 9360

cctttgcttt gtgtatgtac cctacatgtg gttacatcct ttcatctaca agcgaagcaa 9420

aacagcaaaa aataaaacag gttttagaat atggagcttc ccttctaagc cttcaaacta 9480

tttagcttct gtatgcgtgt gcttgagagg gaaggttttt catgttccct ttcctgtccc 9540

ctaactccca cagcttattc agaggagacc aacttcaccg atggaagtac tgacatgaca 9600

agagcttcag aagatacttc attttcacct tgattcaaaa caaagacttt ttttttgtgc 9660

ctaattttac atttcatgga gtcacgatgt ccagatttct tattacaagt tatttgttca 9720

ttacagtcct cccctaatat taaagcctca acacgtagca ttcctcaacg tgaatcccag 9780

agagactcca gatgtcaagt tggtaaaatt taacgttgat agagcaaaca aggggtccaa 9840

actagcagct ctgcaggagt tgcatggaaa cttcctatgg cacaacattt tgaatgtcca 9900

gatttgactg caataattaa ggactttgcc ttaatttgtc acagtgaatg gacgcatgca 9960

aatacagctt attgccagct aatacatata ataataatta aaaaaaaaag actgcaaaaa 10020

gtagcaattc atcatttttt aaaaagaaat tgtaacagaa attatgtcca gtatcacctg 10080

aattcaaaac aattacacca tagctctgaa gtagtatttt agatgtattt acctgtgagc 10140

tcccttttta tattaggaag attagctgga cacgagttgc tgatgtttag tcctggaggc 10200

ggcatgcttt ggttgccata aaggtcaatc aaattcccag acacaggcaa ctagaaaaga 10260

caaggaagga gaaagatcag ttggtttgcc ttatgaaaaa gtttgtacag aacaaatgag 10320

ttatcacctg ctaggaagtc tgtgctcttt tccatgtgca aacagagctt gcagtaaggt 10380

cagcaaaaat tatgcatatg catgggcatg aggaataaac cacctacact ggttaagttc 10440

catcctctct tctcaggaag gtgcagacaa tactttatta ttattttttt gaaatgcaag 10500

gaagcacagc tccatttaag ttgaaaaaaa ttaagaaaga aaaggtaatg cacactgaca 10560

agtattgctg tattgttcct cacaattgct ctgccgaaag ctaagatctg taccctcagc 10620

aattccaagc aaaaaagcgc ctgaaagcgc ctacagaaga ctcatggagg atgcccccat 10680

ttttccaata acacgagtta aagttcttag caaatcatta caagcttgtc agaggacact 10740

gtattcatca atcacatttt tcatgtgatt tgaaggctag ttacagagag aacgttttca 10800

taagtttaac ttgtttttca ccttgcgtaa caaaaatatg caccggttgg aggaagactg 10860

ttttgacata catgtcctca attactcatt cattctgtta ttcttcttta ggtatgttta 10920

agcaaagaga ttcataagga aatagagtag tattccaaca agacttagca aagcaatccc 10980

ttaacattac aacgtaaatt taaagaaatt cccgtgtatc ctgagtggat tattttacct 11040

gcctattctt tgttttcaaa ctaggttttg agaagacagg tataaagtac tttgatacct 11100

tatctgagta gcaacatttc ttatctttgc tgcaaagaaa accagtccct ccaagacact 11160

gatcctgcag agattagacc attaacttca cacccacgta aataacgctg gggaacacaa 11220

tggagtgagt cacagcacgg gctattaaac tcctactgac agtccttgcc agactgagac 11280

aggagaaaat ttctctgcta cctagagcta ccataagaaa cgaagctgat aatgcacata 11340

ttgaagtgcc accacctaga cgccatgctg agatttgaca ggagacttct gaactgacag 11400

aggccaacag aaaaaggagg tcctctgact tggtcttcaa gtgacagagg aaaaaggacc 11460

tcaattctca gcaaatctag tcctacaatt tcctgcacac cacttcgctt ctgaatttca 11520

tatcaaaata tatattaaaa aaatagtaaa tctgttttag ttcccagctg tcctccacag 11580

agtgtgatcc tagttgcaac acacccacat taatgtcact gagaccatta tctcactgct 11640

ctacaaagcc gaccctcaga tcttttggcc cctcgacgca tccgtgacac aggagcagca 11700

gtggcgtcac acaaccttct gacttggacg tgctctttct attcatgctc ccaagtatgg 11760

tgagagcaca aaggacttgt ttgtaggcct gggacttgaa gttcagaacc agacatgtga 11820

aatctaatta ggaatttcag aggaaaaata aaaagatacc agaaactggc atctcaaatc 11880

ctgggtaaag cagctaggtt gtaacaaagc atggagaacg gtcgaggtca ccaagctgct 11940

ttaattattg caagataaac cctgcatgtt acgaaaggac tgtggattgt gattccacct 12000

ccctgtccac agatgtcaac atcatgttgg atttccatgt aaggttccgt aaggaacagt 12060

gcaatcacca caaaagggca ggggctgctg actgtgcaca ccgagtcctg caggaatggc 12120

ttgcaccaga agaacccacc tgagagctgg gcaacaaagc acaagcaatg ctgatgcatc 12180

actaactaaa gaaagatgaa gggagaaatt tgcatatctg attatctttt aaagtttgtt 12240

tctgtatttt aaatgttttc cttaaggctg tttgccgatt ttagctaaga catttaatac 12300

agcatctttc agatcatccc caagaaatta attcacctta actctctcta aaagtaaatt 12360

aagtaagagc agcttatgcc aggcctggtt tgtttccttc cttgagacag aaaggtgtgt 12420

ttttgttcac gagatttcat atcatgaaat atgatgggct ctaataaaat aagcactcac 12480

attcaggcca ttcaggctca gtgtgtatca ccgaggattc tgttccccag aaggtgctgt 12540

gctgctccct tacctacaag ggcacgctca ggggcagtcc ctgcaataag actcttctac 12600

tgctggtgtt cagcttccat tctgacacct tccagcacag cagcaaagct ggttaacttg 12660

tgacccagta ccaaagttta caaacatgtc cataggccct gtattgcaat gagccctttt 12720

caattaggag gtacttagaa acacccctcc gcttgttcta cactcaaatc tctgattttg 12780

tttatcaatt aagtactgtt accaaacgta acaaactggt cctttcattc tttgttggtt 12840

acggaacaat agcagccttt gttgtagtct cagcacagct atttataggt tgcgtctctt 12900

tagaaaccat ttcctctctt tatttttttt tccagctcta cttagctcag tctacagaca 12960

tcaattcctt attagtaaaa tactctgtgt tttcagagct aaatctcatt ccaaatgaaa 13020

agtgtttgtt atggcaggct cgtttttccc caaaccaatt gttgaggaac gtgtcttgtt 13080

ttccatcact tcacgactgg taccaggtta attcagaggc tgaaacatgc tttttgcatc 13140

gagcagttta atggtattag tagactgctc caagggcact atttaagggg ggaggttggt 13200

gggctgtttt ccgcagcatg caccatgcct ccaagagggc tgtgggaatc tgtgggagcc 13260

agactcggtt acatgacatc agacaagtac tgtcagcaaa tgtaaaccta tgctacaaaa 13320

atcgatgtta agtggtaagc tgcaaatttt ggcactagat aaaaaagaac gagacgccag 13380

cactggcaga catctactca gccagatcta acagcactta agcctaaggc aaaaaattca 13440

ctagagctac cctccacaaa agatcaaaat aagattatgc cagaagtacg attatgttaa 13500

tgctcgcagc aatgtgaacc agcaaacaga agtggcaact tggaggtaat gcgtgccttc 13560

ccttgctcac caactttccc atctcagcat tcttgtcctt tgtcccagac ttcctgggcg 13620

cccttctccc cattcagttt taagctttat tctttcacct ctggtttgct ttgaggggtt 13680

gctttttttc ctttcttttt tctaattttg taccatcttt gttgtcatct aacttgggac 13740

ttatctccat ttgcaatcag actaaatata tgatattaga gcttcacaat taaattagag 13800

cagctttggg cagggatgct aacacttaaa tgtggcttac agtttcttat ttgcacataa 13860

taagatggga acctggggtg ccaaaggttc cctctaggtt tccaaagcca agcttgatgc 13920

ccattaatat acccattaat atcaatgggt ataagcttac ttctgttagt tcaattcact 13980

cttctactat aatcttccca ctgattaacc tgaatttgat ttaaaaaaaa gcaaaattgt 14040

cctatgtatt ccctcaacct gactccaact tgcatttttg aactcacgtt cttcactgca 14100

attgtatact ttatgatctt caaagaaaag ctcattattc tccacagata gtgcgctaac 14160

tgcttaactg ctcagagaaa aaaaaaagtt acactccagt tgcacattca cttgaggagc 14220

agcgattagg cttaatgggg aaagcaaaag gaagggacca ttgtaccgtg tttgccatct 14280

gcaaggctgg gtccatcaag ccaaggattt cttcattgta gcttgattct agactaatta 14340

tgtcatcgat cacatcgtcc atctgcaata gtaaaggtga acaataaaaa atagaacagt 14400

tatcaaaacc aagaaggaac aaaggtctaa caaagcactt cagcaacaca ctggttctct 14460

gcagtatggg ccaaagcaga ggtctgcaca attctgccat actccaaatc taatccccct 14520

gacataactc tccctatcca gctagggctg tcagaccaca acaaagcttg aggcttcatg 14580

atctgttcca cacctcccat cagcagttac ctgcctacgg acagctgttc ctcccaggaa 14640

cagcagagga tatcggtgcc cgtacaacaa gcctcactaa ggatgtggaa ccctctacct 14700

tgttctgcac ctggtcccac taccttctca tctccaaaca cagggctcca gcactgcctg 14760

cccaccttcc accttgatca tccaggtgtg gggcaaaaag tgacttacag actgtaacgc 14820

ctgagccctg gcatctgcag agggaaggag aaggcttgct gctgaaaggc tgccctccct 14880

caacggggaa cctggtttat ttcctaatac tccatcccag gaagcaccac ccaagagtag 14940

ctcagtgatg attctaggtg gtgcaaacag tttttggggc tgggctggag ggttcctcca 15000

taagttccta catggtcaag gtgcaagaca gtcttccaaa gcactgctta gatgctctga 15060

gctgacctgc agcttgcacc tatttctttt tggatcacta taaaaaaata agcttaagtg 15120

acactaatca aaaataataa taatttcaaa aaaaacgtga aattgatagt acaaagagta 15180

gtttagggga ttttcatatc agcgcaataa agtatgatca gaatgaatta cactggctga 15240

aagtgagggc aagaaaggaa acaatagtta cagcccatat ctgaagtctt gttactaaaa 15300

taaaatgcag tctgatcaaa ataataataa taaaaaatac aaataattta aaaaatgact 15360

actgaggaca aaattaccat ggatggatta aagccttctc tctctacctc ccccttaagt 15420

gcaggtgtga tctgcctaaa aatgggctct tagaaactgt ctcagcagca acacgtacat 15480

gcagcttggg ggaaaaactg ctttaaaaac gacttcctga agtgagatcc taaaacaccc 15540

ttgataattt cttctgctat agtgttagag ttcttgttac agttctgaaa agggttaaca 15600

cttttctaga tgttttaatg cacttgatat caatggtaat atgcaaggac atggcagatt 15660

tgtaatgtaa gtgggggtta tgttgtttca tttcttaaat acattttaca ttcaacttgt 15720

taaatacatt actggtcaag tttactattt agcataagtt caaagaaaca ttctcattgg 15780

aaaggtgttc agtgctccaa ctcctgtgtc atccagtacc tgcatgcatg atgctcgaga 15840

gtgtgtattc agagccgggc actcactctc aaccctgctt tgctcttcaa atttataaaa 15900

tccctgcaga aataaaaagg aaaaaaaaaa agcaaagcat ggatctgagt tcatggcaca 15960

gtctcagtcc caaaaagagc acgattgaaa cagtgtcatg cacacatgtg cctgatgaca 16020

cagcaaagtg aagctggaaa taacttgaga aaaaaactgt ggtacaagtg agatgatagg 16080

atggaacttg gattttcact taatttaaaa ctatggagca caagtaattc ctcttgcatg 16140

tgtaaaagcc atttttaaat aactgttttt ggagttggat aaatttttcc tcacagagtg 16200

catctgtatt aaaattctgg cacaaaggct cagaggaagg atggggaaca aagggagctc 16260

tataattctc tcttgtttgc caagttgcta aaatattgac atgattcttt tacttgctaa 16320

tgtctttcaa ataaccttta ttttgcacct cagcacatac aacatccaac aaagtttttg 16380

ccagcaaaga tttaagacaa gattgtgaac gacttctagg catagaagcc tcaaatctgc 16440

agcgcagaaa cagcaaattg gaaacatttc aggatatact agctcagata atctggtgtt 16500

gaaactaagc aagagacttg ctcaaatttt agtccaaata tttctaaaac cctaaaatat 16560

ctaaaaaaaa aaagtctgtt gtttttcagt tatttttgtt atgctctttg ctctttctac 16620

actgaatggg ccataaataa cattattcca gaaagctaca gctaacaaaa ctgctttgta 16680

ttctccaaag gtcaactatt cactgcgtgc atcaagcaag caaaaaaaaa aatagaagtt 16740

ttaataagca taaactagct aaaatagttc aaaaaaggaa attacatgac agaggggttt 16800

ttttgtttgt tttttgccct aatgttttat acaaatacat attcatcagg caggcaaaga 16860

agcattaatt gcattttcaa actgagcagt tcagaaaaga agataaacgc aacatcttaa 16920

ttagcaaata tagtttgaaa cggaaagaaa gcagaatgca attctgtttc actgaagagt 16980

atgcacaaca agcgtgtatt acctcttttt cacagttgga gttaagcgtg agcatggcca 17040

tcgggctgtt gggcgcgctg cttccagttc caggcggcat gacatgatca ccgggctggt 17100

ttggacatgg caagctcagg gcttggttgg cgtgtttatt tgctagagtg gtagagaggt 17160

actgctttac ctgctgccgc tgggcttgct ggatatggta cttggttggg ttctcaaggt 17220

gagtctgcac ctgcatgaca ggagttagga aagggcttta ttgttttaca acgtcattac 17280

gtgtggcatc gatctgagct atacatctgt gacacagacc agctaagcat ttgactcaaa 17340

gttacaaacc cgtgcggctc caaggaactc aacagggctt cactaattca cgttcagtgg 17400

caaggaatga gttcctgaga gaatgttcca ataactgact tattaacagg aaagcaacga 17460

gttccacttc tcttccacat ggtgagattt cccacttaaa atcaatgctc cagttttcca 17520

gttcacacag acactctgca acaggtactt ctctgttcag accttcagca acacagtgtt 17580

taattttcac caaaatatgg agaaataagt gccacagcag gactcgtttt acttgtcaat 17640

agctcctaat aaaagctaaa tagtatcaac tgctatcctc tatcagattt caagtgaagg 17700

acagatttaa tgatttgttg ttggtttttc ttttgctttt taaaatatca aatgtgttat 17760

taaaaaaaaa aaaaaagtaa tgtcccattt tttgtctcta gatgatttat aactcagata 17820

tatggcctaa gtcacctaca aataagacag catgtctaaa aaaacacgtt acacaaagaa 17880

gatgcaagaa aagcagcgtc tccagtcaga tcaatcactg tcctcaatag cagccatttt 17940

tgtatttaag ataacctttc cagcacttca aaatattttc acatataaaa tatgaaaaaa 18000

agcacaccaa gctattctct tccactcgaa ataagcacca gaagaaaagt tttttttgtt 18060

tttttttttt ttaatggaaa aagtcaccat atcttggaaa tgccgtcacc tttctttgat 18120

cattaatatc tttatcaaaa ggtagaaaac aaacaagtaa aataatactc tgaaaagttt 18180

gtttgttttg aacagttcca gcaaagtgac ttctattacc gttacaactg accgtgcacg 18240

ttccccactc atttctgtgg aagaaattaa agcaaactag atacaagagg aaaaggattt 18300

tctgtccact gtaagccttc ttagcaagag cttttaaaat aagtctgcaa ataactcacc 18360

tgataatgat tatactcaag catttccagc ataatactta taggttgtga gagaagtgac 18420

tgcaatatcc tttgctttat gctactccag tctgagcgat tcacatatgg aagcaccata 18480

aacaagctgc cccaaaccag cagcagaaac tggaacacca ctgctcagac ttatagtaac 18540

ctccgctaat tagctacgca tgtagagaaa caaacagttc acacaggggc tgtgcaagtt 18600

tagccctaag tattggtatt ttttccctgg ctttgacgtc atgctttttc ataaacataa 18660

aaggaccttt taagtgatgg cctatcaaag tcagattgac agtttgctca aggctaactc 18720

gctattcatc tttcgttcca gtcatactga tagacaatgg tatttttctt tagcagtaag 18780

gttaaaagtt tttaagtacg gaggaagaaa taaccaaacg aatgccctac gagcccaacg 18840

cacatccacc agccatgggg cgctggatga gcccgcggcg ctatgcagcc ctggtgcagc 18900

cgggagccac agcagcccat ggccctgccc ttggcacccg ctgctcaggg cgccagctgc 18960

acccaggacc tgcagctcca cagggaggat ttgaggattt caaggagccg tgctgcttct 19020

tcagcctcct ttgtgcccat tcagtctgtc ccagctttca gcagcaaggc ttccagtgta 19080

ggaagggttg acaatgggct aaagcacatt gcaaatatac ctctgtccac ccaacaccat 19140

tactgcacac agtccctaca cctcccagac ctttcttatc cttgctactc aatcgcttga 19200

cagaagtgtt gtgctgaagc acatgcatct gaagttgctc tacttgtgct gttttccaaa 19260

gtctgctgca cagagaaaca tacttcagat agggtgttaa gagagcttaa cctgatttgg 19320

tgcaccaagt cccaacacag tccggcagcc tgctgcgcta agtgcagcac ttgctttgtg 19380

agtcccaagc ggtgttatcg attttcatat caccagtggt aagagttctg ctacccagat 19440

aaacctattt ttcacttaaa ttttcatctt ccaagtagat gaagttatca ttaaatgagt 19500

tcactttcgt aacccttatt ttaaaaacaa tcttacagta ttcgcacagt ccttttcctt 19560

ccccagcatc aaaaaccact catgctccct cagccagaat gagggacgga actaagcctc 19620

ctccctgcat caaccacatc aaccacagat ccatgctccc aggagaagga caagcaaaat 19680

tccaggatcc acatctccac tggcacaaaa ctgctcccca ggctcgctgt gcaagtgctc 19740

tgcgctgtgc cacaggtccc tgcttcgccc agtgcacaca cgcatgcctt ggcctcttcc 19800

ttctcactgc taaggtcagg catcaagact tctgtattag aaatggctaa agagtaaatc 19860

agcagtaaca aagcttacaa tgaaaggcaa agttgggatc ccgcacactg cctgatggaa 19920

cagatccgtg ctgatgtatt tgtctgccgt ttgcctgtaa ggactcccgg ccctattgtc 19980

tgaacctgca ggctgatgat acaagagctt gcaaaatgtg caagtatcat cacaatttac 20040

agaggtacta ataccccgaa ctgactccac gacacagaaa gttttgaaac acatgctgcc 20100

tacctgacat cagactcttg ttggctaaac aaacactaga aaacaaaaca tcttaccact 20160

attttttttt gtcttatttt gctactccat aatacttttt ttctttaata ggctgtgccg 20220

gcctttgttg cagggaaggc aagaaaaaaa aatgtgaaat tcagcttcag aaagaacaca 20280

gctcctttta aattccagca gacattaggg tatcatttaa taacagcatg acaaataaaa 20340

tcctgtgtct aagtagacag aagaagagca acaagagcat gaaaatgcag cgtttttact 20400

tcagctggaa catatttggc agccaactgg cacttctgac aggactttta gtcagaaact 20460

aggaagtaag gctggacatt tagcttctgt gttaggcttt tactgcctaa cagccctgca 20520

gagccctctc aagaatcgca tcaacctttt ccacctctgc ctctgaagtt atttcagttc 20580

tcttatatca gatttctacc aattacagaa gtgtaactat tctctgatcg ttttaacctt 20640

actgacaagt gtgttctctt tggcacaggg caacagactt caccaagagt ccagaggcta 20700

tttgggtgac gggaattcct cctcttcaca gggaacataa ctgctagaca aacagcactg 20760

aagatttaca tgcagcagaa ctgcaaagac aaattgtagt ctagctggtt taattcctaa 20820

cacaccacac agtcaccaca gtcactttgg ttcagaaaca acggcacagt cttgtggaag 20880

tattttgaac aaactttttt ttttttaatc ttgttataca ctggtaacaa ctagctctga 20940

ggaaatgcat tttctgtcag gtgtctggac ttcaaataag cagcagagaa gactattgct 21000

tttacagaga aaactatcag aagagctgtg gtcattttct gctagttgta cgcagttctc 21060

cagggccatg ccaccttctc acagaagtcc agtagtgaac tgcagaacac agaactgcct 21120

tttctgtcat taaattgctc tgaaatgatg caaaggggag atttggtgag agaacatttc 21180

ctctcagtct gtagctgatc caacatactt ccgtgtttga ctaaaagaac gaatgattaa 21240

tctcagaacg agggggcctg caacactggt attccagctc ctcacagcag tgcaacaaaa 21300

ccagcagatc ccacaggatg gaaagcatct ccacacggtt caggcagata tcacacctcc 21360

accactctta gtggtggggc acacaccgag ctgtgcctgc accgaagccc agcagcacaa 21420

caccccgtgc ttccctccta ggccatggtc aggcctagtt cttcaagcag gagctagcgt 21480

atggcgagag cacaggccct gctggtcaga acagcttacc ccggtttcct cctccaattt 21540

gataggaatg tatcccccaa gaggatacag cacccacttc tgaggaaaaa gcggattatt 21600

aaccctccaa acaaggagtt ggccagcaaa gctgaagagg ccccagtcct caattgtggt 21660

ggtacatctg caagagtctc ctgataacct atcatgatgc cgacagctct agtcccccac 21720

aggcagggaa ggagcacatt tgagttgcct gaatcacaca gaagattctt ctcatcaaaa 21780

aacaccactg aacctttctg tgaaagcaca gccaaaccac ttaatgcttc atttatgcag 21840

agacaggctt gggcacagga acataccgaa gatttagaga ggacagctaa ccaaagtggg 21900

aggaagaatt tttcccctca cccccacatt tttttttatt tttatttttt ttttaattaa 21960

aaagaccatt tgtttcaagt aaactgcctt aggacttaca gtttataatt acaaataaaa 22020

tacttttcgg tcggttttct cagttcaata gaactttttg ttgcttagac agctaaatgt 22080

catgggattt ttaagaaagg ttaaaattct gcgtcgttta acaattaatg gtacccaaag 22140

aaaaattata taaaaccaca tccagtgcca tgttttgcaa gagctacttt gctggcttca 22200

gttaggttcc ctgcaccagg aagccagctc agtatgctag cccaggcctt cacagctagt 22260

cttgttttta aagctgcttc aacccatcac tttcagggat tagcagcaag ttgttttcta 22320

ggtccttgaa tgaagtcaaa aacctgtaca aacaggtgtt tatcatttca catggggtaa 22380

agtctgagaa caactaaaag gaaagacaat caattactgc ctcttatatc aagatgactg 22440

cattttactg tcaaagcaca tatacattgt taacttgatt ttgagcagct ggcaaattag 22500

tcttaaacat aaataaccat tcaaagggga ttcttgtagt gtgagttatc attattaccc 22560

agccttaaaa gcctttattc tccctaggct ttctaatcca aattcaatat tctgcaaaca 22620

gtattcatca cagactggcc tattatgcct agaaatctga agcaccacat ataattccaa 22680

taattccaat tatttcatta aaatattgat catgggtttc actgcaatgc caggaagaac 22740

cctcacagta tccaaatcag agaaatttaa atatccagga agacaatggt ttaactcatt 22800

ctcaaagaca ttttcccaca tgcagcattc aaatgttaag cactgacatc acatttttcc 22860

ccttaaggac atctgtggtc tgaagtagct tcacattgta agttcagtgt tcttgctttc 22920

ttaaaccttt tcagtcctcc aggatgaaac agttacctca cttgaaatat aatcaaaaag 22980

aagttgattg ggttgcagta cgtatcattc ccaagggtag ctgggaattc agaaacagat 23040

atacctattt cataacttct caggctgttg ttctcccctt ctctccatct ttgctagagc 23100

atatttgctc cctctctttt agtccataac ccaggggcag tgactagaac aacactgacc 23160

agaaagccca gggagaaagc tgaaaagtta cttttactga ctggggaatg atgtgcaaag 23220

aagaatttat ttcagtattt tatcttagga catagccaaa gcaaggaaat cacatctgtt 23280

aacacttaat aaaactctct ggttacttac ttcatccaaa acaccctctg aacatgaaaa 23340

tcaaggcagg tacaagaaag ccatttaaga gtcagcagta tgattgcagt ctaagtggtt 23400

ccaacagtca tcttcaaaca cagttatgaa aaggcacatt gtacagaaag aacatcgttt 23460

acacaactga gaattgtttt gcccatcacc aaaaactcct ctgtgtaacg tgacattttc 23520

ctccaaggga aagtcatgga agcagacaaa aaaaagaaca aagtttttgt gattggtatt 23580

taaaaattgt gctttagcta caggagcaca tgatcagagt atctaaagaa attaaagaga 23640

ggttaaccaa tttgcttaag atgacaggaa ttggtgtaat ttacaggtta gtggcctgct 23700

agctgccatt ccgtaagtgt ggttttcgga cagaatctca tcctttgaac tgctatggtt 23760

ttctgttagg aaataaacat ttcagctgat tattttctga catacatgcc ataatttctc 23820

agttttttat agttactggg taggtaccag aaaccttttc aacatttcag gacagacaag 23880

taattgactc catgaaatgc cagacacaaa tctttgctac gatttcacct ataccagtca 23940

taaatatttt atgactggca gacaacacta ttggccacaa aactgtatct aaccagcaag 24000

cttgaattac tgccaaggta aaaaaaaaaa aacttgattc aagcacctca aatctacatc 24060

ctcctttctc atctgcacta gaggccttga cactgctcac attagggcat gaaagagaag 24120

actctcccat acaggaactg tcaatgacag acataaagct gctccataag atctctcgta 24180

agctggcaga gacacgtcag ctccagagga agcagaattg taggatcaga catctcagtt 24240

gccccaaggc ctgtactgat ggtctgaaac cacttctgtt cccagcttct ccttggaaag 24300

cctttcatag gaaaagagct atcatccata cttcctacag cagcaccaac ttaagattta 24360

tctagaccac tctaacacac ttcgctattg acttgaaaga gtcacttcag attggtaaaa 24420

atatttgtcc aaccttatct tcctcccaag gctgccaagc acggcattag ctagtgccag 24480

ctttgacctg cagtcacata tttagagcct gagcaagcct atcagttaca tttcacagga 24540

gttaccaatt ccgccaccct tccagcactc agtaacccct gctagacctg cacaacagaa 24600

catttccctt ccaggtccct ggttcactgc acctctgctg taaaacctgg tggcttcgaa 24660

cagccttcat tctacagcat ttctgatggc aagaaagaaa tgcagaacag caacactcat 24720

tctcaggagg gccttcacag gtcagcttac gacttaaagt acatcaactt tcacacgtac 24780

aaagggttta gagatttggg atggctttct agagcccaca gctgggagag catgtactta 24840

gctgtgcaaa aggaggaaca tgtatgtcct caaatactgc agggctctga ggcagtttcc 24900

acaggtgaga agaggcacct ttctccaggc cctggtttgc tagggtcact cactgtcaat 24960

cagctgccaa acatgacaga ttttaatcca gtaaatctgc tttgtccctg tatgggcttg 25020

tcttcacatg tatcatgcaa cactgacttt tcacaagtgg aatttaagct aaaactggcc 25080

attttgctca acctgtagat gaaagcaagc aaaacaaacc aaacaaacat tattttcata 25140

acgtttaaaa ctccctaccc tccacaccgg tgttgtcact acaaggtttg tgaaagcttg 25200

ctacatttta tagcatatga gatgaagtta gagcccagat aactgctggt gtagaggaac 25260

actatcttca gcacttgggg gccaagcaag gactaaggct gatgtcagat cgttcaaatt 25320

ctcagctcta gcttgtaaga aagaagtggt gttcagaata gtgcctgtgg gccatccaag 25380

tatttactta caaaatctca acattcagcc aaggagtgca agataggaag agatttaaag 25440

atggcactag atcaaaatat attgaacagc caaaagccac aaacagaacc atacttgtgt 25500

ccttcccttt tttgaacaga agttatttgt aagtgcaaat acaactccta gattattata 25560

ttcaagagga aaaaacctac tttggagagg cacactgcgc accaacagga catccacact 25620

tgttgcatac atatgttaaa aatgatgcag gtgaaatact gctacactct ttgttcacat 25680

aatccacggg gactgtgaga ctggcagcat gctccctcgg tccctgacaa aacacatacc 25740

aagacatgcc tctgctcgta agtgaagtct gaaaagagaa aacacgaacg ctaaacccaa 25800

cgaacaccac agtgcatgct gtgtttggcc tgtaccacta actggaagac cttcctacca 25860

tagcttttaa aggaatgccc aaaaataaaa gtcataattt tgaaaggacc atttatttta 25920

atgaatcact tagcactaca ctggctacag cacaaaggac acttgtttaa gtacgcgcct 25980

gtttggaggc tgttttctga cggacacgta gcgccatgag aaaaaattag caacagaatg 26040

aagatcttgt taattcaaaa cagttttatc cattaaaatg ccctgagctt ctaaatgtca 26100

cagttaaaaa aaaaattaga gacactttgc tgggctctat aatattttgc ctacagccta 26160

cgggtgtctc atgtcagccc caataggaaa ggtctggaga agcagaacaa agcagtccag 26220

atgcacccgt tttatccaca ctgctaatat gatcattaga aacagcctaa aataatacta 26280

cactgctgag ttcttttaag tgaaggaaat gcaagctggc ttaccaccga actgcttgat 26340

ttttatcctt ctgttcaacg agctgtgatg ttatttatga ctgtattaaa acctactgag 26400

ttgtagtaag gcagtttcag tacctcgcag cgctcgatca tcgtgctcac taatgtaaga 26460

caacccaagt caccttcatg caaactcaac cagctacaat ctgaagcatt ctgctaaata 26520

ccagaattgg gattttgttt ttggtacttt ggcaagccac aaaggtgcgt acagccctta 26580

cacagaggca agcctccgcc ctggaggaga attaacccag ccagccgaat tgatgccaca 26640

gagcatcaaa actgagaact gtgctgtcag cattactgct gacaggtgat ggcttttcct 26700

agcaagcaga gtgcttatgt ccctgacact tccattaagc aacattacat attttatttc 26760

ctgacatctc actcacaaag aaggtgttag agcagacatc acagtagcca caagagatta 26820

aaggtagcag cagtaagaaa ttaagtatgt ttattgcact catcttaaac cacaaggaaa 26880

tttagtgttt tagttgtttc ttggctacag gaatgctgcc ctaacacttt tgtaacacca 26940

agatgtattt aaggcaaaga gccataatgt tgtgtgacct cacctttaca ccccaggatc 27000

ttattcccgt aacagccaga tggtggctat cttctggctc caataaggaa aaaaaaaaaa 27060

aaaaaaaaaa aaaaaggcaa aagcacacac aagcttatcg taggctcatc tgtcaaagaa 27120

agatggcaac atttcagccc tttaaagatc tcaagcccta catacaccaa gatacagcat 27180

cacgatctat tttataccga caaaaacaac atccagccac tcggccactg ctgtaacctt 27240

cggcacacct gcccctccgt gtctcgcaga tgagctgttg aaaacccttc tcctcatgct 27300

atttcacata tggcaggcaa ataatcagtt acactgaaac aagaagcacc tgtccaccta 27360

aatgaccctt gacaggccag aagatggagc atttgcagca ccatcacctg aactggggca 27420

agctttccac acaaattcag tcagctgtgg ttggctctgc tcagccagcc gtaaccaaaa 27480

aacgcagtca ggatcattac ttccaagaga tgttgcacaa gcaattttta tttttatttt 27540

ttattttttt ttaaatcatg aatacttaaa agagaggctg tgaaaaaaat gctatatgaa 27600

aagctgaccc aatcttctct tttccagcac aaacccatgc cctggtagca gcagcctttt 27660

cagcactgac tctgtacaag agcaagggcc tgtgagctct cgatgctctg ctggctgcag 27720

tcctactgac acagcagggt taggactctc ccgacgggac cctgaatcgc attttgtggt 27780

gtggcagcac actaattcca ccttccgaaa cattttacag aaagctcttt tcaggggttt 27840

catctgaact tcaaaaaagc acaagtcaac agctcaccag gagaggcact tcacgagaca 27900

caaagccggg tgtgtgttat catccccatt ctccaaaggt tagctgcaga tcagtattgc 27960

caagaaattc agaagtattg gttttgtcag aagatcaaaa cgtctcttgc gagagacatt 28020

tagggcctcc aggcttggtc tcagaagaat cctattttaa accacactga tgtaaccctt 28080

caattgctgc tacaagaacg gaactaggtt cccaagggga aacaggcaca gcactttcca 28140

gcacagcctt actcgtgggg aaaatacaca caaagcctct tgctgctgga aaacaggcaa 28200

tgctgcatgt gtatcacgac acctgtcacc actgatggat agcgtacaca aagtttcagc 28260

ttcctaagag agtatggaag cataacccag ctccaggcaa gccgagtctg gacaattaga 28320

tggaggtaga gattagatgt ggagtaacta gatcctaact caatctcccc accttcctta 28380

ccccttttta tagctcaagc ataatcccat tttagcaaat cattatttta catgatggaa 28440

tgaacagctt gagatcttaa tgctttgaaa atgcaaatag gcacacaccg aactgtgttg 28500

gattttaaag gaaagtttaa atggaggaac aaagcaaggg agctggaggc agcagaccta 28560

atgacagcca gcatcgaggc ccccccatct cggtgtgtgc ccaccagcag tgtgctgcag 28620

gacacggggt gccaccaggg actggctgca gacaccacgt atgccccagg ccaccgctgc 28680

cagcccggct caggaccgat ctcacaggct gagttctcca tagtggctgc agagcagctt 28740

cttgtttaat ttcctctgac atgaatgtaa ttagtactca tcaacagcac tctgcgaaaa 28800

taaactcact tgtaataagt ggggacaacg gggaattgac ttccaaagca cgttgctgct 28860

ccacgctcca ggctgctggc caccagggct gcaaggaaca ctctcagcca cctctgctgc 28920

aaacagtcca tgtaacacag aactgaaccc cccaaaaacg ggtgtaaata agcaataggt 28980

tgaataatag cctgttttgc ttctgcctta ccccatctgg tggcctactg cttatgtaca 29040

gctggaagca acagttaatt gaggcgcgtt atcgggaatt gcagaactag agaaataaac 29100

gtggcactgg atgagctgcg gtgccctaac agttctctgc tgctgttccc tgagtgccat 29160

ggcagtcgca ctcctcactg catcccaggt ggagcatctc gatgccgtgc actcacacca 29220

cgtggagcca tcgggaccgg gcgccgtcac cgcagccccg cgagctggtc gtggcggcct 29280

caaccgaagc ggataagtgg aattaatacc aactcctgga gaaggtggaa actcagaaag 29340

actttctaag gatgaggctt gggagctgcc tgactcaggg agaagccttg gctggaagct 29400

gtcccctcgt tccttacatg aaatgtcgca gagagaggca gaaggcagct ctgagcaacg 29460

cacatgggaa gcatcgcttc gcacagagca cccacttaat ttaataagaa gcactccaga 29520

gcagaaggca ccacttttct tccctcaaaa agccaatata cattagaaaa gcataaattc 29580

ttttgaaaag ccacagtagc atgattaaat actatttttc cttttacaca acaaacctta 29640

cagggctgca cccttaagga atatatgaac tttcatgtga aaggagccca ttaactgctg 29700

cacaatgaat ggtgcctgtg gtccaagatt catttccttt tagaaagcag aaaggacgga 29760

gagcttacaa ctgctgcaca caactgctga ctcattcaag gtcactttac ttcttccttg 29820

aacaaactgt ttctttgaga cccagcctat cgtggtattt aagactgtag tttcaagagg 29880

gagtcaggga acacaatttt gttaggctgt gccaaatttg caggacccaa tattatttca 29940

aatttaagtc cgattttttt ttttcttaaa gtactattca gaaacgaaaa tccattttgg 30000

cagggtgggg agaggggatt aaatttcaca gaggtcactt taaacaagcc tacaagtact 30060

tccaaagccc ttaaaagtag attttataca gatttgtgtc atgtcagcat ggctatcaaa 30120

cagatttcaa gagacataat ttccataaaa cagtcagtag tactaggtta gttctagaaa 30180

tagtactaga ttatgtaaat aaagactttc aattattgag ttacgataga gatgactggc 30240

gtgtttttta aacttaagtc tcacttctaa gaatttgctg tttaacaaga caagaacgaa 30300

cttcaaagcc ctttctgaaa ggacagcgag caccacatgg cttcagcaac acagccccag 30360

atcgcacggg gctgcagcag cagtaagaga agctctgccc cactggcttt gaacagaggc 30420

tttccacttg ctacctgctc atctcttcct ctaaaaatca gccatgaagt ctgtaaagtt 30480

ttgatggttt cagctaggaa tttccaggtg acttttcgga agaaagcctg aaacagaacc 30540

tttgatttat ccaccgtgac acacaagctt cctaacaaat tcctagctca cttctgattc 30600

ggagtcagaa taatttctgc tttttccacc acaagtgcac tgagcacagc agtcgactgc 30660

aaggccttta actaggaaac aaaaaatgtc agtgttagat agggagggga aaaaacactc 30720

ccaaaatcac ggtgaggatt ttctagagct tacagcaatg actccaagtc ctttaaagat 30780

gcttgggtgc ccagcaccat ctgagacccc aagccctcct agcaaggcca cacagctccc 30840

tcccgatagc ctatgatgct ttttggggag cagatctgcc tctccccatg ttcagaagga 30900

aaagagagat agatacgaat acagtgcccc gggacagaaa aaatactctt taaaacacgg 30960

ctttttgttt gtgtttttta aacacaagac gcatttctcc tcatttcagt gtgcttgttt 31020

ttggatacat tgcagattcc aattttgatt ttctttattt aaagcattcc agctgcactt 31080

gctgaacaag tttactactt ccacttgctg ccagtagcag gtttactttc aagagagagc 31140

ctgccttcca cagattttct agcagcaatt tgtttccaca tgtaggaacc gttgtgtagg 31200

gcgcacaacc ggttaatggc ttaaagacac ctctgggaca tgagttggtg gaaaccaaat 31260

acaatataaa caagacaatt agaatcatgt gtgaaaatca gtttctatca cttgactgtt 31320

agccaccgga gaagggaacg gtctggaaag gcagcttcag cagcaactga gagctttccc 31380

aagaaataga cgctcttgga aaccataatg ccatctgctg agagattatg atagtgcaat 31440

gcaaggaatg acagccctga attagttctc cagtgcttac agcatttcat ctgctagctt 31500

aattactgta atatcgatgt tgaaaggcat gctgcttctg gtcatgttca aaaataaaaa 31560

gaaattgttt aaatgggata agttatccaa gggttttcaa agaactacct gagacatcac 31620

agcctactga taatcagcaa gtcaaggaca ctaaatcaca gaggtccata attgaaatgt 31680

tgttccagta tgctgaaagg atacaagtag gatgtgggac acacatattc atttggctgg 31740

gacactctaa cattgcaaaa acactgggaa ttaaatagct ctttttgatc tattatgttt 31800

ttaaagttac tctattttgt gctccgtatc tttacaagct tgagaatttg tacataattc 31860

taattcaact acaaaaatat atcaccaagt ctcaaaatta caaataggtt aagtgtgaaa 31920

ctggaatcca atagaagtgc cagcgacagc ctgcaggacc gagctgttac cccaatatcc 31980

caccagcact ctccatagaa atgtttggag gcagcaaggt atttttgtag ttggtagcgt 32040

agttgctgtg aactaaagat tggcaggata atgattccac aaagaccacc agatggatta 32100

tttcatttcc caatctgcca agcatctaaa atcaggccct gttttgctgg gtcacaacaa 32160

aacttaatag aacatgagga acaagaacca gccattggga tgcaaagggg atgcacgcat 32220

gacaagccag gaggatgttt ttccttattt gcactaaacc aagtctgccc ctgaaggtac 32280

tgatgaatta caagaggttc agaaagcctt ctgaaggatt caagatcagg cacgccccag 32340

attcaagatc taagcagaac actagcacag tctgcttttc ctctagccca caagtttcag 32400

gagtgttgca atctgagcct gtcattaatt attaaaggaa gatttacagt aataataatt 32460

taattctcta agtcagacag acagaactaa ccagatccac tggctagtaa ctgatggtgt 32520

atgaaatcct aagtggatgc tgccgcacag aactgttctt ccagagaagt gcaaaggcag 32580

cagcctggtg atccagttct agtctgaccc tctcatttca gttctgagga atgcctctcc 32640

tatgtgtaaa gaaagggaaa aaagattttt ctataatatt ttaactgaag aatgctgcat 32700

gagtgtcttc ctaaggaaac acaaaccaaa tgtgttccat cctcagctca gggacaattt 32760

ccactgcaag cagtacagag ctgcattact aaaatggatc cttttggtca aagaacaaga 32820

catggggctg gcacctatct gtgaggaata aagagttaaa acagctacca cttcctcaac 32880

tggcccactg ctattgtgag ggcagtgttc agctgaattc catcaggcca cgctgtctta 32940

acataacctg acgtgttcct cttcaagatc tcggaatttt tttctcctct ctgcttttca 33000

gcagcataca ctagtaaata aatacacccc aaaagccaaa tggtaatcca agtaaaaata 33060

atatttaatt atatagtggg gggtagtgag gggactgagt aacttttttt tttttttttt 33120

ttttttaaag taaaagactc gccaggtcct tcacagacaa cacaatactg gtttttgttt 33180

caagttcata acttggctct taaagtagtt gataatgaac cagcaggcat tattttaaat 33240

caaccagttt tattaaatga gccagaaagg aaagaggcag ggctaattac cagcattcct 33300

tgatgaatta cctagcggtg tgtggtaaat ctctgggcgt ctttgtcttt atccccaaat 33360

ttatccctta cttatttctc aagaggttaa aggacacggg gctaattaag gcaccaatcc 33420

aatgacagac agaatcattc aaggtaaggc tggtttttac tctttcaagc ttttgcatct 33480

ctgcctggta cttcttaatc taatttcttt tttttttttc cccccttaat ataactgctg 33540

gggaggggga ggaaaagcgc taatcttgtc acggtcaggc aggttgaaat tgccatgcag 33600

tgctgggcaa aatgctgcat atttgtaatt tagccaataa gaaattcctt aagcctctcc 33660

tgaatggcag gaatgtctct agcttgtcac ttccaattca gtcactgcgc aggttcggcg 33720

tctcgcagaa taatgctgcc aaactagctg aggaactgaa atatttgcga gtgtacataa 33780

tgcctaaaga ttactttaag cagatctgag ttttcacatt tcaacattta acgtctgtca 33840

aagctgtcga cagaggcctc cccagacctc cacatgactc cccccagcct tcagcgccgt 33900

ggggggctcc aggctgtgct gcagaggcaa ccagcacaac cccaggcacc agcactgccc 33960

ctttctgtgt tcttggctca accggtgcag ggtggtaagg tggttttatt ttaagctgac 34020

ttgggctgaa acaggatttg atacaaccta ggcaacacac acactgacac ccatagcaaa 34080

aggagatcgg ggatgaaggg tgaaagcagg ctttgcactc gcagtcaagc agtcagacac 34140

ttgtgcacag aagtacattg cccccttgaa ttccacgcat gtttaacgtc taataacctc 34200

tgtgttgagc aagagtttac attttacctc tagcactcac cacagacatc agcaccagtt 34260

ctcattctgg aggagcttac aacccactaa gcaagcaaaa gactgaaggc ctgagagata 34320

tcttacttac gtgaaggctt aggacaggct ttagaggttc tgtggttaaa agcaagtttc 34380

tccttctgaa gaatttgctg tgggctgaag ttacactcac cgactctgtg ttgtatctgc 34440

agttcagcaa gacctggtct ggcaaatcac ttctccgagt tacaactgca aatacatgcc 34500

aggctggctt aacagactgg cacacaccgc tagctgctat tgtcaggttc gattaaatta 34560

acttttgatc agataagcat ttccacaaac acacatttac tgtaacagat gctaccacac 34620

ctttatctcg acatcccgtt gttcctgttc tacagaactg taggacagat agattgatct 34680

cccatacaat gaagtctcgt ggtaattatt acaccaggaa tactatttct aactgcacag 34740

ccttccctgg tgagcagaag gactacatta acaccacagg tcgctctttg taacagagca 34800

cgctggaaaa ggtctgggag gaggcatttt ttgatacttg tccatagtca ttaacatcac 34860

tcagaggcat gcttggcatt tcaaaaccga agaaatacag ggtatctgat gcaaagagct 34920

tatcatggag gcacatttaa aacctgaatg aacaatgaca cgagggttaa agcaagaggc 34980

cagccgggcc tcctgcattg tcccagacag cccggctgcg cttctgttga ctcaggaaaa 35040

gctgctggaa ggagaaagat aagcgtgagc tacagcagta ccggggtggc agcaagatgg 35100

gtgaatggac ccactctggg atcagtggcg atcagcctgc aacactgcca taggatgggc 35160

aaagaaggac aaagcaagaa tggaaatgca gtgaacacca gttggttctg gactttgaaa 35220

agcagatatt tcagcttgtt gggggcccca ggcagaagaa aacctgggat ggatgtctgc 35280

tgcccactgg tgttttcaat cctgcctgtc ctggtgatgc tttcaaagga attactctgc 35340

agttctgcag gccgaggatg ctcgcagaca ccagggtcag cagacacagt aggactacag 35400

gtgtaaaaat gattctgtga tatcaagaag acattaacag atgaaatttg aaacagctgt 35460

gggaaaactt tttaacagac cggcttccct ccaaacctag ggtggattct cttgatttct 35520

gatagccaag gatttcacca cctgaaacaa actgagctct gcaaggatta acagcagcac 35580

tgttctgcag atgaatgacc tgggagcagg actagcctct catcactcct cacaggtgag 35640

gggcagaggc ctgatgccca acctcctcca gcaccgggct gaccccgctt acagaaatgt 35700

ggggatctgc tcccagaggc cagccctgca gcagccagca ccgagagctg cagaggtgcc 35760

tctgctccag caggaaagga atcctaccgg acaggaggat aaacaacagc aacaacttct 35820

tgctatcaag gacacaaaat tatcatacac ctgtatggaa gaaactattt tataaagaaa 35880

accactgaag atattttaag aagaattctt tacaaaagaa gttcatttct taaataaaaa 35940

gtattgactg actgtgtaca ttgggctcct gatcaaacat cagcacattt taaagttact 36000

ttctactatt tctgaggcag agcaagaagt tgaagcagag cagaagcaaa gaacctgctg 36060

tgacacctcc acgtagcatt aaccctaacc agcagcagga gatggatgag aagatgcttg 36120

aagacaaact agcccttacg tctgtgaaac taacatcagt ggagcagaat gacaactgaa 36180

acccctaata aaccctcctg ctattagatg agaagagatg ctgcttgata tgcattagcc 36240

tcattcctag tccatcacaa aattaggctt cactaagcaa cactacattt taattccaac 36300

aggtcatttc ctaccttgtc aagctacatt acttgacata gaaatgatgc aataagaagt 36360

cattactcaa gaaaataaaa tcctatttat tcatataccc ttgaatgtaa tggccacgtt 36420

atgtaacttg gaaattccta tgcaatgtga catggattaa tacgtattac tctttctgta 36480

ctgtttctaa atgacatttt aaacatctcg aacacaaata caaaatgcaa actcatcaac 36540

agtttcagct tctttttaag ctagtcattc caaactacta tcctcagaca ctcaactact 36600

actcccctaa tatcgggaga tacgggacac agcaagccac actgttaatt acataatcca 36660

aagagtgaac cgaatcaaca ggttgctttt tctttcctct cttttttttt tttttttaaa 36720

tgtgatttta aattctataa aaatcatcaa aactaacaac aaagcagctg ccacttttgg 36780

tcttatttgt gtagtaacat tcaaaaagct gttttaaaga aaattgcctt gcagattaac 36840

atttctcaaa gtgtaaaaat cccctttttg tccacctcat tctttaaggt cactaaatcc 36900

agaccatctg ggtaataaag ttaccatgtg tagttcccta ttcacctaat taaaacttct 36960

gtacatctcg tatggataat ttgtctatag ctaaaaccaa ctgaagtgta aaaagcagct 37020

ccggggaaat gatgtcacta gaagcaggtt actgctggca gagctggctg tcagcatatc 37080

ggcaccagga agatcccaat ttactgttaa acacatcttt tgagccaagc aacctatttc 37140

ataataataa taaatcatat ggactttcca gtaaaatcct attcacactg aaacatcaat 37200

agtaaatatt ttaatgttaa aaaaccacac agcttcgagc ctggtattta gcactatttt 37260

tgtacagagt tggtccagtc tggtagcact acggataaaa gcacgaatct ttagctgctt 37320

ccactaaaaa cataagccgc aaaagggagc aaaccattct ctcttctctc aacccgagca 37380

agagaccaag aggactgggt tttgctagtt ttggctgtta cttatgtaaa ggcagataac 37440

gaggagtcag acgccgtttt gcgtgtttcc gcagccatta cctaaaacac acgactctgt 37500

tgccaaaaag cccttgctgc cagctattac ttttcattta caagccaggt aaggatgcac 37560

tcacacagct ttttgctctg tggggtcaaa cattctgccg aggactgggg acagatgctt 37620

cctctgtggt cccaaaggac aaggattcct gtttgatacc ttcacgtgca gttgctagca 37680

atgctcccgc aagttacgga tctgcaggaa tccctgtgag aagccagttc ctaggcactg 37740

cgcttgtaat aacagagcat cacctttcaa cacagcatct tctgtaacct gtctcctaga 37800

aataaagggc tgccttccct tctctctcac gtatggaggt acttgtaaaa aggacactac 37860

agaaaactgc taatgcttca agagattaca catcctcctt gatagtatgg tatcatcaga 37920

aaataaaagc tttcctgtaa gtttacgaac agatataaaa aaacaacact acaatttatg 37980

tcagacagta ttaggatgaa agtatagaaa aaactaacac atgcttggtt aaccaagtgc 38040

actacaaaca gaaaactact ttgtcttcta cacatctgct agatccaaat tgtaactaca 38100

gtgtcataat ttggaaatac atctaacaaa ttcaggatgg acaagcgtta ggataaaatt 38160

gcttttccca gtcaatttct actcctacac tgtagaccat gcttgagtat attctagtat 38220

ccccattgag tattaatttt ggcactgtcc acacaatacc ataatgcagg ttcaataaat 38280

agaaggaccc tacatttccc ccataaaaca gtcatgctga aactgtcttg aaatactgca 38340

tctgcaagcc tatttaaagg ctaaatcttg gaatacttat cgatatggag cagtgatggg 38400

caaagtacaa gaatgcattg taagcaagtt caattaactt ctctggcaag aatataatta 38460

ctaacagaaa gcaattgcca aatatccaac taaaatagaa ggtctagcac taatgcagag 38520

caaataacta acttctcaaa acttacaaaa actttactct aacgaaatat gtctattttt 38580

acatcacttt tcccattatt ttgctaacac attttcacgt tactttatgt ggacttaccc 38640

tgtgcatgaa attctctttt cttctagttg aagaagtttt tccctgagta attagcaagc 38700

tacatcccct ttgacagaga gatactaaca agctcctaca gtttccagta agacaaggaa 38760

tccaaagcta ctaatagcaa acaagtcatt taagcaaaaa aggattacaa aggcttgttg 38820

caagagcctg gaaactcacc catatttcag attaaggctt tctgaggagc tcttttgtaa 38880

tgagttttga ctcaaatttt ttaaaatatg catttttttt aatgaaatat tgacttaaac 38940

atgtattaat cattttttct ttggacaaac aataagtgcc agaaagaccg gacaatgatt 39000

tccaatggga aaactagaga aatggtggtt ttataacaaa acctccctcg agatctcaaa 39060

gtataaagaa aaaaaatcac aaactaagaa ggatttctac tctctggtac aaggacacca 39120

gaataagctg caaatcaagc cacagtgcag cttatttgcg agacttcatt cccaaaagta 39180

cctgtcaccc catccctagg gcacatgaag tttatttcaa tgagagctac gacacagagc 39240

ggcagagccg ccaacgtacc ttgaggacct ccatgggtac ctgcgtggcg gggggcaggc 39300

tggcggggac gctgacgttg atggcaggtg tctggctgac gggcactctc tgctgcatga 39360

actgggctgc ttgctgcttc tgctgctgct cccgacgctc ctgctcctgc atctgctcac 39420

gcatgagctg ctgccgtagc aggattcgtg aggtcat 39457

<210> 2

<211> 22

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 2

tccccattct ccaaaggtta gc 22

<210> 3

<211> 22

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 3

tccccgttgt ccccacttat ta 22

Claims (2)

1. 一种采用SNP分子标记技术鉴别润州凤头白鸭黑色喙的方法，其特征是，所述分子标记位于润州凤头鸭基因组的11号染色体上，如SEQ ID NO.1所示；所述分子标记位于第28397位点，突变碱基为A或T；鉴别时，包括以下步骤：

步骤1）、提取鸭的基因组DNA；

步骤2）、针对MITF-28397设计特异性引物SEQ ID NO.2和SEQ ID NO.3进行PCR扩增：

步骤3）、将步骤2）得到的PCR产物电泳检测，然后测定基因序列，通过基因测序序列和峰图判定鸭喙色基因型；

若基因型为AA，表示受测个体为黑色喙纯合野生型基因型；若基因型为TT，表示受测个体为黄色喙纯合基因型。

2.根据权利要求1所述的方法，其特征是，步骤1）中待测鸭个体生物检材的基因组DNA为鸭翅下静脉血样基因组DNA。