MEGA510构建系统进化树

1、Escherichia coli O157:H7str. EC4045相关基因的系统进化树构建 以 glutamate-ammonia ligase adenylyltransferase 为例 1、进入NCBI主页2、选择Protien数据库，输入搜索名称3、在右侧的Top Organisms Tree下，选择其中一个种4、点击进入第一条目，5、点击send to保存文件6、回到NCBI,至U Blast,页面7、点击进入 Protien Blast ,8、将以上搜索的蛋白质序列输入到搜索框中，进行序列比对NIB Dldbl；yi|Fei 1A DescriptionsDH mi - lij

2、-火 LHJU ,Legend for links to otheresources E UniGene S GEO S Gene S Structure 任Map Viewer SI PubChem BioAssaySequences producing significant alignments:AccessionDescriptionMax scoreTotal scoreOuerv coveraaeE valueMax identLinks |NP 289628.1bifunctional qlutamine-synthetase adenylyltransferase/deaden

3、yltran*19321932100%0.0100%ZP 02822650.1qlutamate-ammonia liqase adenylyltransferase Escherichia coli 015719291929100%0.099%ZP 02810932.1qlutamate-ammonia liqase adenylyltransferase Escherichia coli 015719291929100%0.099%ZP 13639065.1qlutamate-ammonia liqase adenylyltransferase family protein Escher1

4、9271927100%0.099%ZP 17966105.1qlutamate-ammonia-liqase adenylyltransferase Escherichia coli TW119261926100%0.099%ZP 11875205.1bifunctional qlutamine-synthetase adenylyltransferase/deadenyltran*19253850100%0.099%YP 003501250.1bifunctional qlutamine-synthetase adenylyltransferase/deadenyltran*19233846

5、100%0.099%ZP 03048216.1qlutamate-ammonia liqase adenylyltransferase Escherichia coli El 1019141914100%0.099%YP 002409453.1bifunctional qlutamine-synthetase adenylyltransferase/deadenyltran*19131913100%0.099%EYP 006170150.1qlutamate-ammonia-liqase adenylyltransferase Escherichia coli P 1X1X3 1912100%

6、0.099%ZP 07147234.1Glutamate- -ammonia-liqase adenylyltransferase Escherichia col deadenyltrans 详细1912100%0.099%ZP 10059820.1qlutamate-ammonia-liqase adenylyltransferase Escherichia sp. 4_ 没有汉英互译结果3824100%0.099%YP 001459844.1bifunctional qlutamine-synthetase adenylyltransferase/deadenyltn 话尝试网页搜索191

7、2100%0.099%ZP 08344882.1Glutamate- -ammonia-liqase adenylyltransferase Escherichia coli H1911910100%0.099%YP 003000634.1qlutamine synthetase deadenylylatinq enzyme Escherichia coli BL21i19103821100%0.099%ZP 02999084.1qlutamate-ammonia liqase adenylyltransferase Escherichia coli 536319101910100%0.099

8、%ZP 12021155.1qlutamate-ammonia-liqase adenylyltransferase Escherichia coli JE19091909100%0.099%丫P 002404424bifunctional qlutamine-synthetase adenylyltransferase/deadenyltrans19091909100%0.099%NP 417525.1.fused deadenylyltransferase/adenylyltransferase for qlutamine synth19091909100%0.099%1 ZP 08379

9、801.1FGli itAmAtp- - Ammonia- linApl arlanvIvltTAnqfaraqaFseharichiA rnli H19081908100%0.099%9、在上图中选取10个与原蛋白序列相关性最强的蛋白，一一如上述方法保存，以便接下来 进行系统进化树的构建。10、将10个序列导入MEGA，得到11、点击 Alignment，鼠 M5 iTAIignmenCExplorer (sequence 1 .tasta)Data Edit SearchAlignmentAlign by ClustalWWeb Sequencer Display HelpW对 Align

10、 By Muscle昭其略XX|勉督I 41 I*Protein Sequences 如兴让与FAhBirv1. gi1168797643 ItJ Mark/Unmark Site+L Align Marked SitesUnmark All SitesCtrl+MCtrl+L一;TmTnonia. liga我J gi|168785925|t 3. gi|168785925|e 4 gi | H9Q93783 | eIM Delete Gap-Only Sites4 Auto-Fill Gaps一;TmTnonia. IzLga一 FimTiEiiia. ligmE一;TTiummia. Iz

11、Lgms5. cii I 4245Z1991 I refl EP 1796610S.11 .lut amat e - amiLLoni a-1 i .asgi | 416777554 | rmf | EP 118752CI5.il bi funutzLDnm】 glutaiainEi-sgi | 291Z&4432 | irmf |TP DQ35D1Z5CL J. | Li furLUtintiEL】 glutaiiLinE：-8- gi 1193067248|irmf|ZP D3D48216-11 g】uaiiLm*-；=iTrE2rL：La. ligmsw9 gi IZ187D1824 I

12、 Ef I YP 口。24口“53.11 Li funBz.MnM glu七斗心官 1CL g:L | 386706303 I irmf |TP 00617。15。- J. | gLuiz aiiimii EimiTncmim-IzLg:Q Q Q Q Q Q Q Q Q GIVIV IV IVIV vv vvite# F12、出现下图，点击OK/ with L w/o Gaps13、得到序列匹配后的，将其保存为“.meg ”格式EEEEEEEEEEEEEEEEEE-F斜m5: Alignment Explorer (sequencel.fasta)Data Edit Search Align

13、ment Web Sequencer Display HelpD Q=IkATCGw对|G电盖略X K帷i B41 lProtein Sequences如已EL m s /瓦hb m * l-fr * 士 * *, *,* * * *,!* - *, -A- 1_gi |L68797E43 | re； f |07872650 _11glut aniEdz e-FiTiEicinia.liga耳eW _gi IJ.687859Z5 I ire： f IP(JZ81CI93Z 一J. Iglu!iaiiiEdzE-；=iTiEinrL：LmJ:LgmsE3 一gi |J.68785925 | re

14、： f |口Z81D932 一11glut anisJz E-FnTEicjrLia.liga耳e4-gi |4J.9D93783 | re： f |EP13639D65 _11glutaiiisJLm-FimTriDriia.J:Lga三e5 一gi | 4N4 S艺 19911 ire： f |P17966105.11glutanimii.E-FiTiEinnim-ligmse6一gi I4167775E4 | tEf |11875ZD5 _ 11Ei furuTtiunEL】 glutaniinEf-x一 4。|291284437|上-已 IYP CICI35CIJ.25CL J. I

15、Li furu 二七:LnnEd. glut am One： 一gi | 3.9306724:8 | re： f |D3CI48216.11glut aiiisJLE-FnTEicjrLia. liga耳e-g:L | W187D18W4 | rmf |YPDDZ4CI94.E；3 _ 1 | 匕i furLt.iDnml gluiz.aniinEi-J.CL gi | 3867D63CI3 | ire： f | YP DD6J.7D15CL J. | gl】;itaiiLELiz E；-FiTnTcti:Lm-J.:Lgwssssssssss w一 官LLLLLLLLLYllllllllll

16、eeeeeeeeee I TR I I I I I V V 一一o一 QaQo.dQeeeeeeeeeeSite # 14司 (* with L w/o Gaps14M5 AlignmentExplorer (sequent el.fasta)DataEdit Search Alignment Web Seguencer Display HelpD Create New 芽 OpenReopen Closes昭其曜XX|蝠鳄|4I I*Phylogeneti匚 Analysis=5 CL J. |aiaa七 m 一 Pimm urti a. 1 :L gms e耐 Save SessionCt

17、rl+SExport AlignmentDNA Sequencesy Protein Sequencesuf Translate/Untranslate面 Select Geneti匚 Code TableMEGA Formatiiiionia 1 i as *monia ligmswFASTA formatPAUP FormatiDiQnia-liias*glutaiiiine-ial CTlur. snine -16.11 glutaiiiate-aiaiiionia li?ase9453.11 bi fi.uict i on al glut aiiiiiie -?0150.1 | glu

18、t aia ate-a to th oni a-ligs93 2 一 11 glut aiiiar.日 一 ；=nnm cmi a. 1 i gms eHHHHHHHHHvffiAllIiLM一一RlllllllllLLilnu- nnnlL L L L L L LLLLLLLLLLvvvvvvvvvIIIIIIVVVTrE-rArArArArE-rE-1rAMJ Reverse CcimplementFeverseComplementExit Alignment Explorerw/o Gaps15、重新回到MEGA主页面，打开保存的格式文件16、点击TA可看到17、点击C可看到保守序列，V

19、则是可变序列18、回到主页面，点击Phylogeny19、点击下图中Compute20、得到系统进化树跚锅5: TreeExplorer (C:Documents and Settingsfy桌面、新建文件夹(2)sequencel.meg)BnFileCGN郁旧ec备枝痞废的MnImage Subtree View Compute Ancestors Caption Help昭怎|嗟瘴莲胃Original tree Bootstrap consensus tree Igi|168785925|reflZP 02810932.11 glutamate-ammonia ligase adenyl

20、yltransferase Escherichia coli O157:H7 str. EC869gi|168785925|reflZP 02810932.11 glutamate-ammonia ligase adenylyltransferase Escherichia coli O157:H7 str. EC869(2) gi|419093783|reflZP 13639065.11 glutamate-ammonia ligase adenylyltransferase family protein Escherichia coli DEC4C giI4245219911reflZP 17966105.11 glutamate-ammonia-ligase adenylyltransferase Escherichia coli TW14301gi|168797643|reflZP 02822650.11 g