36 7 Vol.36No ActaEcolog iaeanimalisdomastici Jul.2015 氥 氥 欍欍欍欍欍欍欍欍欍欍氥 氥 10 犛犜犚! 1, 2, 2, 2, 1 (1., ;2., ) [" #] PCR! " # $ % & '
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- Orsolya Pásztorné
- 5 évvel ezelőtt
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1 36 7 Vol.36No ActaEcolog iaeanimalisdomastici Jul.2015 氥 氥 欍欍欍欍欍欍欍欍欍欍氥 氥 10 犛犜犚! 1, 2, 2, 2, 1 (1., ;2., ) [" #] PCR! " # $ % & ' ( BM1818 BM1824 CSSM66 TGLA53 ETH10 TGLA126 INRA037 ILSTS006 ILST0013 ETH225 ) 10 * STR +,-./01, 10*+,-.+,23(Pi) 456(H) /0789 :(PIC)) ; :68 (. 10 * < = * ) > +,,A B C 10.2; (D10* STR +,-.AB/0789:C0.685,10*+,-.AB/0789 :E 0.5 D10*+,-.AB 456 AB 456 C F G645HI,JKLMNO.10* STR +,- (JK/ 1 JK PQJKRS. T)UV [$%&] STR +,-; (;JK/01 [' ] S811.6 [ ()*] A [ +, ] (2015) !"#$%&' "# %$95% [1],( )"#$*+,-./0+1,234567$89:; $1 < = >? (microsatelite),@ A B C D E, 2~6 FBGH IJ$ DE [2 3] =>?-BKL89, MNO$( $ -,PQRS<TUV WX:;Y $, NB Z$[\ [4 5], ]^_\/+`:;ab8c 8c + [6 8] "# G =>? [\/"#` :;$ N [9], Y P \ #$=>? 8c"#$ ` :;ab 8 [10 13] HY \ E! (FAO)! :; (ISAG)E $\/#:;8c$ 9 Holstein #=>?G $< =>?G X68 "# H$:;( 8 c, "# G $:;( /0 68 " # H,\ EDTA #- 10 PCRbufer dntp Marker F /,, ;O HEALFORCE ;PCR (My CycleTM ThermalCycler)! BTO RAD ,W DNA XY \ TIANampBlood DNA Kit ( ( ) ), -, PCR Q PCR [ :PCR B25μL,2 PCR Master 12.5μL, 0 0.6μL,DNA 1 μl, 10.5μL, \ PCR 0 STR G $ B:94 10min, 30s,72 45s,! 35~40 " #,$ 72 5 min,4 PCR % & 2% $ ', ()G * G 8+, =>?J,G +-./% <=HIZ[\ \ 10 =>?G \/"#=>?$G,M 9 G (ETH10 ETH225 BM1818 BM1824 IL [ ] : [G45 ] 6 75(2013 N A5) [-8 9] :; (1990-),,,, <= >?Y E mail: @qq.com A B(1969-),C,,Y,(,Y D: >?
2 7 :;, :"#10 犛犜犚 G :;( Y 11 STS006 CSSM66 HEL9 TGLA53 TGLA126) 0E! (FAO)! : ; (ISAG)E $\/#:;8c$ Holstein #=>?G,E < G (ILST0013) 0 $=>?,., F 10X,GH FAM IJ, KL STR Table1 ObjectiveSTRandtheirprimersequences G Locus (5' 3')Primersequence MN TM/ Marker BM1818 ILSTS006 CSSM66 TGLA53 ETH10 TGLA126 INRA037 ILSTS006 ILST0013 ETH225 F:AGCTGGGAATATAACCAAAGG R:AGTGCTTTCAAGGTCCATGC F:TGTCTGTATTTCTGCTGTGG R:ACACGGAAGCGATCTAACG F:ACACAAATCCTTTCTGCCAGCTGA R:AATTTAATGCACTGAGGAGCTTGG F:GCTTTCAGAAATAGTTTGCATTCA R:ATCTTCACATGATATTACAGCAGA F:GTTCAGGACTGGCCCTGCTAACA R:CCTCCAGCCCACTTTCTCTTCTC F:CTAATTTAGAATGAGAGAGGCTTCT R:TTGGTCTCTATTCTCTGAATATTCC F:GATCCTGCTTATATTTAACCAC R:AAAATTCCATGGAGAGAGAAAC F:TGTCTGTATTTCTGCTGTGG R:ACACGGAAGCGATCTAAACG F:CTTGATCCTTATAGAACTGG R:ACACAAAATCAGATCAGTGG F:GATCACCTTGCCACTATTTCCT R:GATCACCTTGCCACTATTTCCT 57 FAM 58 FAM 56 FAM 58 FAM 57 FAM 54 FAM ] P \ Cervus2.0 \G +%0 1, O 9 X H $8c J,$ JG PQ R N ( STU& (PIC) :; [14],VW Hardy Weinberg X V ( PQY +$,` PQ$O$ JG + $1 $ $G R N?Z[\ ]$R N, ^ R N _ $` H$ JG V$PQ 2 a 8c 2.1 :; < \IJ *X=>? PCR % 8 c,a1~3b c8j,$ijs JG $ a INRA037 J, $ JG 8 B bp(a1);IL ST0013J, $ J G 8 B bp(a2);TGLA53J,$ JG 8 B bp(a3) 2.2 = >5?@ 8c68 "# 10 STR G J G % PQ8.L24,TGLA53 J, ] 16 JG,G 151~273bp, J G $ G P Q ~ INRA037J, 14 J G,G 119~134bp, J G $ G P Q ~ ILST0013 J, 11 J G,G B117~138bp M J G 1 INRA037 ABC7DE Fig.1 ElectrophoresessignalaboutamplificationproductsofINRA037site
3 ILST0013 ABC7DE Fig.2 ElectrophoresessignalaboutamplificationproductsofILST0013site 3 TGLA53 ABC7DE Fig.3 ElectrophoresessignalaboutamplificationproductsofTGLA53site $G P Q B ~ ETH225 ETH10 BM1824 CSSM66 J, 10 J G,G 8 B133~150bp 209~236 bp 178~263bp 173~190bp 0 JG $G PQ B ~ ~ ~ ~ BM1818 J, JG B 252~260bp,G P Q B ~0.4722;TGLA126 J, J G B 119~134 bp, J G P Q B ~ ;LSTS006J, J G B 27~ 291bp, JG PQB0.0217~ Hardy Weinberg. \ Cervus2.0X10 =>?J,$ Hardy Weinberg X V [15], a b : BM1824 ILSTS006 b X ( 犘 < 0.01),TGLA53 BM1818 F 2.4 FG HIJ.L 3 4 K,10 = >? J, $ PIC 0.367~0.849, B0.685 ( STU& 0.5, BON(,L G N,:;( ;VW L HY \$=>? P N $:;ST,4 Y"#$:;( "#$ V=>?J, $^ R N B0.7995; R N$ B0.333~0.868, R NB , /ONR ONR +`,NO$=>? R NL P $:;ST&,N O$+`R NL +`$:;( [16] 3 Z 3.1 KL M! =>? Y DNA ab( $89: ; < < ` +$:;( O :;,MX# $ [P, M8 [ $# :;R NY < ` J G _ $,M J G _ F R 9 $ Q [17] R N $ 4 Y : ; a b N$O,R N O ` :;, :;( ; ` $:;, :;( 3 Jurg [18] ( J, 7 B : ; R N B 0.10 $J, [19] X! 6 +8cL :;R N ~ c $ 10 J,, R N(MeanHe)B0.6154, / "#$ R N(0.800) [20],"#+`:;( N, "# $,G"#+`$ N,-$<, 3.2 (N! O =&:; $3 (
4 7 :;, :"#10 犛犜犚 G :;( Y STR?@ Table2 Alelefrequencyof10STRLociinyak G Locus JG Alele PQ Frequency G Locus JG Alele PQ Frequency ETH ETH BM BM ILST CSSM ILSTS TGLA TGLA INRA
5 14 36 J, Locus Table3 ^ R N H(O) F HIJG Hardy Weinberg Heterozygosity,polymorphisminformationcontentandHardy Weinbergof10STRLoci R N H(E) ( STU& PIC Q(1) Excl(1) Q(2) Excl(2) :; X HW TGLA ND TGLA NS CSSM NS ETH NS ILSTS BM ND BM ILST NS INRA NS ETH NS STU& R N JG %,0 % <7 % O, ` $:; O : [21] \ 7 = >? $ PIC B ; ' [22] X $ ` 8c,M PIC B [23] \5 = >? G X + :;( Y,a L =>? ` :;( HY,10 G $ PIC B0.685,ILSTS006 BM1824B N( G,M BON( G ; J NB , JG %B10.2, 10 =>?J, PYHY "#` $:;(, * O$ N H V Hardy Weinberg X Va b ILSTS006 BM1824 / X,M 1 4P./ $, >O, B -\ F$G + G PQ$ 3.3 G DNA 8 PQ RS = >? J, J G 8 + O \ DNA % 0 DNA $ % 0, ^, N, -, N DNA \ 7 \ G + $,G 7 \ $ Mark % 0,?, N DNA $ a,v W W B ^,V $ % 0 V? V $ a V $ DNA 8 c \ $ 8 <, 4 \ ( J,, \ DNA ` % 0 8 c 4 a Z HY 8c$10 =>?J,, R NB ,( STU&NO,b ] $:;( NO$ M ` J, / X,4P$1 : 9, H 6 "#, ` # N $!" NNO, F #G $P ]] = >? : ; " # $ Y N,\/"#$ J,N, V <7$ $%, & < TU : [1] '4,,,." # 8 9 * + $? Z U ([J]. )*+ :,-,2006,32(1): [2] GilmoreS,PeakalR,RobertsonJ.Shorttandemrepeat(STR) DNA maekersarehypervariableandinformativein Cannabis sative:implicationsforforensicinvestigations[j].forensicsci enceinternational,2003,131(1): [3] A./,A0.=>? M[\[J]. Y,2001,2(4): [4] 1,.,,.= >? DNA 8 c X 23` :;8c $ N[J].! U V,2006, 14(4): [5] 456,7.Hmlh1G ; 9 CpG 8 G,= >? 67 9:[J].! ;,2011,8(7):9 11. [6],,A,. \ = >? Y + $ : ;( [J],[\,2009,20(1): [7] A<=,>?@. \=>? 8 c A B : ; ( + [J], C *,2012,18(5): [8] A D, 6,E J,. \ = >? DNA 8 c! [J].! R,2012,48(13):1 5. [9], F. "#` 4 = >? J, : ; ( 8c[J]. :,-,2014,42(5):
6 7 :;, :"#10 犛犜犚 G :;( Y 15 [10] Nguyen T,GeniniS.Application ofbovine microsatelite markersforgeneticdiversityanalysisofswissyak(poepha gusgrunniens)[j].anim Genet,2005,36(6): [11] QiXB,HanJL,LkhagyaB,etal.Geneticdiversityanddif ferentiationofmongolianandrussianyakpopulationa [J]. AnimBreedGenet,2005,122(2): [12] "# +:;( $=>? 8c[J]. (,2008,16(2): [13] '4,, JB,."# +$8 [J].,2006,26(7): [14] KalinowskiS T,MarshalT C.Revisinghowthecomputer program CERVUSaccommodatesgenotypingerrorincreases successinpaternityassignment[j].molecol,2007,16(5): [15] KLM,NO. 5 =>? $( 8c[J].,2014,36(2): [16] O., F PQ> ` 6 = >? G : ; ( Y [J].,2014,46(3): [17] JurgO.AnalysisofHumanGeneticLinkage[M].Revisede dition.baltimore:johnshopkinsuniversitypress,2001: [18],:,R S,.! 6 +$:;8 G T[J].DU,2007,28(3): [19] :,R S,,.7 = >? DNA ( ) +` $( [J]. :,-,2004,32(12): [20] ',FVW,X.,.5 ` =>?( 8c [J].DU,2012,33(1): [21],Y.F,A!,.\5 =>? 8c +$:; [J].! R,2009,45(3):8 11. 犌犲狀犲狋犻犮犘狅犾狔犿狅狉狆犺犻狊犿狅犳犜犲狀犛犜犚犔狅犮犻犻狀犢犪犽 YANG Qi lin 1,ZHANGjun 2,XUShang rong 2,PENG wei 2,WANGZhi qiang 1 (1. 犙犻狀犵犺犪犻犝狀犻狏犲狉狊犻狋狔, 犡犻狀犻狀犵 , 犆犺犻狀犪 ;2. 犙犻狀犵犺犪犻犃犮犪犱犲犿狔狅犳犃狀犻犿犪犾犛犮犻犲狀犮犲犪狀犱犞犲狋犲狉犻狀犪狉狔犕犲犱犻犮犻狀犲 , 犆犺犻狀犪 ) 犃犫狊狋狉犪犮狋 :Thegeneticpolymorphism distributionsoften STRloci (BM1818,BM1824,CSSM66, TGLA53,ETH10,TGLA126,INRA037,ILSTS006,ILST0013,ETH225)ofyakweredetectedbypoly merasechainreaction (PCR)andfulautomaticcapilaryelectrophoresis,andgenefrequency (Pi),het erozygosity (H),polymorphisminformationcontent(PIC)werecalculatedbyCervus2.0software.The resultsshowedthat102alelicgeneswerefoundamongthe68yaksbythe10 microsateliteloci;average PICoftenSTRlociwas0.7546,andthePIC>0.5.Observedheterozygosityandmeanexpectedheterozy gositywere1,0.7995and0.6154,respectively.theresultsindicatedthattheabovetenstrlocicouldbe usedingeneticdiversity,geneticstructuralanalysisandgeneticmappingofyak. 犓犲狔狑狅狉犱狊 :STRLocus;yak;polymorphism
犘狉犲狆犪狉犪狋犻狅狀狅犳犇犻犪犾犱犲犺狔犱犲犆犪狉犫狅狓狔犿犲狋犺狔犾犆犲犾狌犾狅狊犲 犆狅犾犪犵犲狀犆狅犿狆狅狊犻狋犲犎犲犿狅狊狋犪狋犻犮犕犪狋犲狉犻犪犾
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