大菱鲆_Scophthalmusmaximus_生长性状相关的微卫星标记筛选.doc

405Vol.40,No.5Sep.,2009OCEANOLOGIAETLIMNOLOGIASINICA20099(Scophthalmusmaximus)*1,2111111(1.266071;2.201306),,DNA,30(F)(S)10DNA,7PCR,,1(SmaC10)355bp;4(SmaC02SmaC06SmaC09B11-I12/6/3),258bp182bp226bp175bp226bpSmac09,0.354SmaC09,微卫星标记,大菱鲆,生长性状,分群分离分析法Q953(Scophthalmusmaximus),2005),,Turbot,,,,,,,,,,(,,(,2002)2003),1992(SSR)DNA(RAPD),1999(AFLP),(,(,2002;12,2002),2002),,,DNA(,2006),,(18(,2001;,2004;,2007),*,2006BAD01A12012;,nyhyzx07-046;;“863”,2006AA10A404;,2009-ts-11;“”,E-mail:[e-mail]:,,,E-mail:[e-mail]:2008-04-20,:2008-07-3057840(Carmenetal,2007),,,PCR60()(Cnaanietal,2003)PCR1.3.4PCR2h,,(BulkedSegregantAnalysis,BSA)PCR8%,,3μl,360V,,,(Sambrooketal,2002)PAGE,11.3.5BANDSCAN,1.16300,716cm,SPSS,,90%,(Pearsoncorrelation),,Pearson()()13cm30F(fast);,10cmS(slow),∑nx−xy−y20iir=i1=∑∑()()xy1.2nn22x−xy−y(Chenetal,2007;Iyengaretal,2000)iii=1i=130,1.3.6,1,TADH5,1.3(TAKARA,)1.3.1DNAF,BLASTS30DNA,DNASalah(1997),41.3.7(EPPENDORF)Genefinder,50ng/μl,2020(10),1.3.2BSA(BulkedSegregantAnalysis),,PCRPCRFS10,SPCR,5μlDNAF30DNAPCR(PCR∑4()x2=O−T2/TBIO-RAD,PTC0200)25μlPCR,200μmol/LdNTPs,0.8,iiii=1,100ng2DNA,10pmol2.5mmol/LTaqDNAMg,1PCR,buffer,1U942+2.1BSAPCR,PCR30,4min,9430s,FSPCRPCR50s,727250s,30;,17min(Chenetal,2007),FSPCR1.3.3DNAPCR(1),DNAPCR,DNABANDSCAN5:(Scophthalmusmaximus)579表1大菱鲆的30个微卫星位点的引物序列Tab.1Primersequencesof30microsatellitelociinS.maximusMgCl(mmol/L)(5′3′)Genebank2()SmaC01(TG)SmaC02(AT)SmaC03(CA)SmaC04(AT)SmaC05(GT)SmaC06(GGAT)SmaC07(CA)SmaC08(CT)SmaC09(TC)SmaC10(TGA)SmaC11(GTSmaC12(TA)...(GT)F:TGTTGCTTTGCTCCTTTTCCR:TAGTGGAACGGCGTCTAGGTTGTCTCTCGTCAGTGGCAGTR:AAACTGCAGCCTCCAAGATGTGGACTGGTTTACTGCTGGAR:AGGGAACCGATCTGAGAACAATTTGCTTTGAGCCTGATGGR:TTGCAGCACAGTGGTCTTTCTTCGAATTCACCAAGTGTCCR:GTCAGCCATCATCTCCACCTGACCCAACGAGCACTGTTR:GGGCCAACATCATTATGGTCAGCAGCCAGAGCACCAGAR:GCCTCAGACTACAGGAGGATACF:GTACACTTATTGGTGCAAGGCR:TTCTGACAGATTTGCTGGCACTCGACCCGCCCACTACAGGATR:AGCGACAGCAACGCACCAAGGGTGGCTGGGTAAATCTGTTR:CTTCCCTCCGTCTACGCTCCAACCACTAGCTGGAAATCAGACCR:CTTCCCTTCAACTCGGCAAAACTGCAAGCCTTGA