36 7 Vol.36No.7 20157 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., 810016;2., 810016) [" #] PCR! " # $ % & ' ( BM1818 BM1824 CSSM66 TGLA53 ETH10 TGLA126 INRA037 ILSTS006 ILST0013 ETH225 ) 10 * STR +,-./01, 10*+,-.+,23(Pi) 456(H) /0789 :(PIC)) ; :68 (. 10 * < = - >? @ 102 * ) > +,,A B C 10.2; (D10* STR +,-.AB/0789:C0.685,10*+,-.AB/0789 :E 0.5 D10*+,-.AB 456 AB 456 C0.7995 0.6154 F G645HI,JKLMNO.10* STR +,- (JK/ 1 JK PQJKRS. T)UV [$%&] STR +,-; (;JK/01 [' ] S811.6 [ ()*] A [ +, ] 1005 5228(2015)07 0010 06!"#$%&' "# %$95% [1],( )"#$*+,-./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 $:;( 1 1.1 -./0 68 " # H,\ EDTA -20 1.2 1#- 10 PCRbufer dntp Marker F /,, ;O HEALFORCE ;PCR (My CycleTM ThermalCycler)! BTO RAD 1.2 -. 1.2.1 +,W DNA XY \ TIANampBlood DNA Kit ( ( ) ), -,-20 1.2.2 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 +-./%01 1.2.3 <=HIZ[\ \ 10 =>?G \/"#=>?$G,M 9 G (ETH10 ETH225 BM1818 BM1824 IL [ ] 2014 11 28 23 :2015 03 04 [G45 ] 6 75(2013 N A5) [-8 9] :; (1990-),,,, <= >?Y E mail:2580560727@qq.com [ @-8] A B(1969-),C,,Y,(,Y D: >?
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, KL1 21 34 STR 56789 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 1.2.4 ] 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 B124.06 132.42bp(a1);IL ST0013J, $ J G 8 B 121.53 127.29bp(a2);TGLA53J,$ JG 8 B153.80 155.86bp(a3) 2.2 = >5?@ 8c68 "# 10 STR G J G % PQ8.L24,TGLA53 J, ] 16 JG,G 151~273bp, J G $ G P Q 0.0074~ 0.2426 INRA037J, 14 J G,G 119~134bp, J G $ G P Q 0.0074~0.3971 ILST0013 J, 11 J G,G B117~138bp M J G 1 INRA037 ABC7DE Fig.1 ElectrophoresessignalaboutamplificationproductsofINRA037site
12 36 2 ILST0013 ABC7DE Fig.2 ElectrophoresessignalaboutamplificationproductsofILST0013site 3 TGLA53 ABC7DE Fig.3 ElectrophoresessignalaboutamplificationproductsofTGLA53site $G P Q B 0.0109~0.4485 ETH225 ETH10 BM1824 CSSM66 J, 10 J G,G 8 B133~150bp 209~236 bp 178~263bp 173~190bp 0 JG $G PQ B 0.0075~0.2612 0.0074~0.2941 0.0161~0.5597 0.0100~0.3235 BM1818 J, JG B 252~260bp,G P Q B 0.0278~0.4722;TGLA126 J, J G B 119~134 bp, J G P Q B 0.0100~ 0.3300;LSTS006J, J G B 27~ 291bp, JG PQB0.0217~0.4348 2.3 Hardy Weinberg. \ Cervus2.0X10 =>?J,$ 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 0.6154, /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 0.8208~ 0.9336 8 c $ 10 J,, R N(MeanHe)B0.6154, / "#$ R N(0.800) [20],"#+`:;( N, "# $,G"#+`$ N,-$<, 3.2 (N! O =&:; $3 (
7 :;, :"#10 犛犜犚 G :;( Y 13 22 10 STR?@ Table2 Alelefrequencyof10STRLociinyak G Locus JG Alele PQ Frequency G Locus JG Alele PQ Frequency ETH225 133 0.0075 ETH10 209 0.1691 134 0.2612 212 0.2132 135 0.0149 213 0.2941 136 0.3731 217 0.0074 138 0.0075 220 0.0074 140 0.0075 220 0.11250 142 0.0075 221 0.0956 144 0.1791 224 0.0074 146 0.0149 232 0.0147 150 0.1269 236 0.0074 BM1818 252 0.0556 BM1824 178 0.0161 257 0.1944 179 0.0522 258 0.4722 181 0.1269 259 0.0278 187 0.0299 260 0.0833 191 0.0373 250 0.0448 256 0.5597 257 0.0161 258 0.1269 263 0.0323 ILST0013 117 0.0809 CSSM66 173 0.0368 118 0.0074 174 0.1691 120 0.0147 177 0.0147 121 0.0588 183 0.0100 122 0.0217 184 0.0809 123 0.4485 185 0.0147 125 0.1544 186 0.1471 127 0.1471 187 0.0294 129 0.0588 188 0.3235 131 0.0147 190 0.1397 138 0.0109 ILSTS006 27 0.0217 TGLA126 119 0.0200 271 0.0435 121 0.1700 273 0.4348 122 0.0800 274 0.0217 124 0.3300 275 0.0652 126 0.0100 279 0.3261 127 0.0100 280 0.0652 128 0.1000 291 0.0217 129 0.0300 130 0.0400 132 0.0100 134 0.2000 TGLA53 151 0.0515 INRA037 119 0.0074 152 0.2206 120 0.0074 153 0.0147 121 0.1471 154 0.0588 122 0.0809 155 0.0441 123 0.0515 156 0.1029 124 0.3971 161 0.0221 126 0.0074 163 0.0809 127 0.0221 164 0.2426 128 0.0588 165 0.0147 129 0.0147 166 0.0147 130 0.0368 167 0.0294 132 0.0074 168 0.0515 133 0.0294 172 0.0074 134 0.1324 272 0.0074 273 0.0221
14 36 J, Locus Table3 ^ R N H(O) 23 10 F HIJG Hardy Weinberg Heterozygosity,polymorphisminformationcontentandHardy Weinbergof10STRLoci R N H(E) ( STU& PIC Q(1) Excl(1) Q(2) Excl(2) :; X HW TGLA53 0.868 0.867 0.849 0.423 0.268 ND TGLA126 0.333 0.727 0.749 0.595 0.416 NS CSSM66 0.657 0.847 0.794 0.526 0.352 NS ETH10 0.360 0.766 0.786 0.545 0.370 NS ILSTS006 0.735 0.708 0.457 0.867 0.713 BM1818 0.691 0.788 0.663 0.702 0.522 ND BM1824 0.676 0.852 0.367 0.739 0.552 ILST0013 0.735 0.824 0.713 0.639 0.454 NS INRA037 0.338 0.811 0.768 0.561 0.381 NS ETH225 0.761 0.805 0.703 0.660 0.482 NS STU& R N JG %,0 % <7 % O, ` $:; O : [21] \ 7 = >? $ PIC B 0.9024; ' [22] X $ ` 8c,M PIC B0.8293 [23] \5 = >? G X + :;( Y,a L =>? ` :;( HY,10 G $ PIC B0.685,ILSTS006 BM1824B N( G,M BON( G ; J NB 0.6154, 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 0.6154,( 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):114 119. [2] GilmoreS,PeakalR,RobertsonJ.Shorttandemrepeat(STR) DNA maekersarehypervariableandinformativein Cannabis sative:implicationsforforensicinvestigations[j].forensicsci enceinternational,2003,131(1):65 74. [3] A./,A0.=>? M[\[J]. Y,2001,2(4): 315 320. [4] 1,.,,.= >? DNA 8 c X 23` :;8c $ N[J].! U V,2006, 14(4):276 279. [5] 456,7.Hmlh1G ; 9 CpG 8 G,= >? 67 9:[J].! ;,2011,8(7):9 11. [6],,A,. \ = >? Y + $ : ;( [J],[\,2009,20(1):190 196. [7] A<=,>?@. \=>? 8 c A B : ; ( + [J], C *,2012,18(5):944 950. [8] A D, 6,E J,. \ = >? DNA 8 c! 2 + + [J].! R,2012,48(13):1 5. [9], F. "#` 4 = >? J, : ; ( 8c[J]. :,-,2014,42(5): 30 31.
7 :;, :"#10 犛犜犚 G :;( Y 15 [10] Nguyen T,GeniniS.Application ofbovine microsatelite markersforgeneticdiversityanalysisofswissyak(poepha gusgrunniens)[j].anim Genet,2005,36(6):484 489. [11] QiXB,HanJL,LkhagyaB,etal.Geneticdiversityanddif ferentiationofmongolianandrussianyakpopulationa [J]. AnimBreedGenet,2005,122(2):117 126. [12] GSH,A @.!I "# +:;( $=>? 8c[J]. (,2008,16(2):156 165. [13] '4,, JB,."# +$8 [J].,2006,26(7):9601 9605. [14] KalinowskiS T,MarshalT C.Revisinghowthecomputer program CERVUSaccommodatesgenotypingerrorincreases successinpaternityassignment[j].molecol,2007,16(5): 1099 1106. [15] KLM,NO. 5 =>? $( 8c[J].,2014,36(2):36 37. [16] O., F PQ> ` 6 = >? G : ; ( Y [J].,2014,46(3):71 72. [17] JurgO.AnalysisofHumanGeneticLinkage[M].Revisede dition.baltimore:johnshopkinsuniversitypress,2001:112 115. [18],:,R S,.! 6 +$:;8 G T[J].DU,2007,28(3):22 26. [19] :,R S,,.7 = >? DNA ( ) +` $( [J]. :,-,2004,32(12):69 74. [20] ',FVW,X.,.5 ` =>?( 8c [J].DU,2012,33(1):39 42. [21],Y.F,A!,.\5 =>? 8c +$:; [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 810016, 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 810003, 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