5
nopÒÓ¡Ý6qr Tyrp Eist;ouvá.w£23 MN ÛOP QRÀ #S Ä ? þ T UVW X#¦ 1. ?÷%&¸;h&# &'*?÷ 225009 2. *.¸;òã#nI;Ó·¿ª3*?÷ 225009 3. À*.¬@AA.$Zµ:À*¬@ 315400 45 +,3YZ[\`@]^ Tyrp T¦Ä_Fû, <'i。[ D5ÔZ[ Tyrp TÊ`,aÊ`b c©,C|7b¹PQdµe2wx PCRSSCP Singlestrandconformationpolymorphismanalysisofpolymerasechainreac tionproducts NÃË Tyrp T CDS Ê`,<H &f®。[ Z[#g µTYRP1 ]^6åh_,i7]^ñ#$ jkñ\ùl4mZno<HËp Tyrp T CDS , 483bp ãq r ,estúq-uß< crÁ,túvñw,7úAUC AUT xy)。HardyWeinberg ÐøËJâÞ@z{ÄZ[õ|ã-}ÐøfH^ÄZ[õ|ã-ÐøfH@z{[õ|]ûf<H#gmg0.25< PIC<0.50 ), ^ÄZ[õ|]~f<H#gmgPIC025 )。[ ZqTyrp Z[,¦Äñ0_,ÕÖrstZ[ì]Z[¦Ä,LoLÝyz0_,ñò678 Z[Tyrp SNPs Ê` µ 9:;<= S829.1  >?@AB A  >CD= 0517-6611 2016 09-164-04 AnalysisofPolymorphismofExonsinTyrosinaseRelatedProtein1Tyrp1 GeneinRexRabbit Oryctolaguscuniculus HAO Ye SHILijuan YANXiaorong WUXinsheng etal 1.CollegeofAnimalScienceandTechnology YangzhouUni versity Yangzhou Jiangsu225009 2.JiangsuKeyLaboratoryofAnimalGenetics&BreedingDesign Yangzhou Jiangsu225009 3.Zhe jiangYuyaoXinnongRabbitIndustryCo. Ltd. Yuyao Zhejiang315400 Abstract Objective Todiscussthemolecularmechanismoftyrosinaserelatedprotein1Tyrp1 geneofRexrabbit Oryctolaguscunicu lus duringtheformationofhaircolor. Method TheTyrp1genesequence thenucleotidesequence andthebasicphysicalandchemical propertiesofcodingproductswereforecastedandanalyzed.Atthesametime PCRSSCP singlestrandconformationalpolymorphism was usedtodetectthedistributionofCDSsequencesofTyrp1. Result BioinformaticsanalysisfoundoutthatTyrp1proteinwasanunstablehy drophilicprotein whichhadasignalpeptideandafunctionaldomainofthetyrosinasefamily.Thepolymorphismdetectionresultindicatedthat theexon2ofTyrp1in29bp thegeneCDS483bp hadaC Tmutation andthenucleotidemutationdidnotleadtotheaminoacidchange AUC AUT isoleucine .HardyWeinbergequilibriumtestshowedthattheChinchillaRexrabbitpopulationwasatHardyWeinbergdise quilibrium thewhiteRexrabbitpopulationwasatHardyWeinbergequilibrium.PICofchinchillaRexrabbitpopulationwasofmediumpoly morphism0.25<PIC <0.50 ), whiteRexrabbitpopulationwasoflowpolymorphismPIC <0.25 Conclusion Tyrpgenehascer taineffectsonthehaircolorofRexrabbit.ThisresearchprovidescertainreferencesfortheselectionofhaircolorofRexrabbit. Keywords Rexrabbit Tyrp SNPs Sequenceanalysis EFGH ãj²³©³|WCARS441 ); KLæ ¼@+PAPD2014134 )。 IJKL MN1992-), _^st¦stDºR HIªÝoE¡¢£¤¥¦§ RHIÝoªstMNOP 20160221 Tyrp @»>B H3CÐ%D}¼@»KPWDED{#FÓH¢£»¤ºFÓH¶¡{ Tyrosinaserelatedprotein1 TYRP1 ×<&¦n (}¼a;CÐDÇHËÌ+FÓH<G}¼ìÊH U S?@HIJÓ\BHL´(}¼DÐ6Zá nÓ¯PX Tyrp DED S?@HIJÓ \BH>FÓHÓ¯Dó¢»#FÓTyrosine TYR )、 F Ó H ¶¡ { Tyrosinaserelatedprotein2 TYRP2 ÃÐFÓH¶¡{-K lZ56} ~Tyrp @»üØTI#(}¼DaÐcMÛL ¾µÞ*FÓHDûBbcI#VÀ(}¼pQD ·ÞBCY(}¼@sDZTUÖ× 3-4 ¥¦¹º PCRMZ Tyrp Hè`D| Tyrp |«¬a;&?NnÞÉ PCR-SSCP UØT4|Dæ6Hè`«¬<¨c34n Tyrp @»OO}P.Oryctolaguscuniculus (QD< ¨cßàP#P.¡}ÐbQP.¡}ã¡ÃµD1 QRST' 1.1 VZQR 1.1.1 ©ª¸;XÞR¢naäD ÃSTd U}P. 392 ô{}P. 174 ôX 566 ôP.³£©ª Í©ªHQOwÀ*.¬@AA.$Zµ:1.1.2 ©Ö{H 10 ×TBE Tris EDTA KHßà_ ©ÖO a; )7 Z µ: 10% º V Ó WTEMED SDS y÷_©ÖO"RXa;Z µ¼½:1.1.3 ;I;89 NCBI ,-):oD. Tyrp X |YÖ}NC_013669.1 ), n0O Hè`8èD¡ U=8I;ÞCоucQD;;Ka7a ;7)Zµ:CÐ;?N} 1.1.4 Tyrp |nêÂÉ ExPASy 7Y@» DED{/c/«¬nÉ ProtParamn{D ¶n/0U_ß·_ËBc/É, )Oa7Y SMART http //smart.emblheidelberg.de/ ), n TYRP1 {·8Þg NPS@5Zå)D SCOPMA nQ4{DS4·É TMPRED http//www.ch. embnet.org/software/TMPRED_form.html «¬{/|D [ n & ProtScale http //web.expasy.org/ protscale/ «¬|D\ÏcnÉ SwissModel http //swissmodel.expasy.org/ TYRP1 DE{«¬Ø abDc þ abde ¶· [\]^_`JournalofAnhuiAgri.Sci.2016 44 ): 164-167 196

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Page 1: nop 1Tyrp1Eist £23 - nykxw.com · :2 uvÆ29 AA´ BBEi{p@smn Fig.2 ThesequencingresultofAAandBBgenotypesatexon2

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 S829.1  >?@AB

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 0517-6611(2016)09-164-04

AnalysisofPolymorphismofExonsinTyrosinaseRelatedProtein1(Tyrp1)GeneinRexRabbit(Oryctolaguscuniculus)HAOYe1,2,SHILijuan1,2,YANXiaorong1,2,WUXinsheng1,2 etal (1.CollegeofAnimalScienceandTechnology,YangzhouUniversity,Yangzhou,Jiangsu225009;2.JiangsuKeyLaboratoryofAnimalGenetics&BreedingDesign,Yangzhou,Jiangsu225009;3.ZhejiangYuyaoXinnongRabbitIndustryCo.,Ltd.,Yuyao,Zhejiang315400)Abstract [Objective]Todiscussthemolecularmechanismoftyrosinaserelatedprotein1(Tyrp1)geneofRexrabbit(Oryctolaguscuniculus)duringtheformationofhaircolor.[Method]TheTyrp1genesequence,thenucleotidesequence,andthebasicphysicalandchemicalpropertiesofcodingproductswereforecastedandanalyzed.Atthesametime,PCRSSCP(singlestrandconformationalpolymorphism)wasusedtodetectthedistributionofCDSsequencesofTyrp1.[Result]BioinformaticsanalysisfoundoutthatTyrp1proteinwasanunstablehydrophilicprotein,whichhadasignalpeptideandafunctionaldomainofthetyrosinasefamily.Thepolymorphismdetectionresultindicatedthattheexon2ofTyrp1in29bp(thegeneCDS483bp)hadaC→Tmutation,andthenucleotidemutationdidnotleadtotheaminoacidchange(AUC→AUT,isoleucine).HardyWeinbergequilibriumtestshowedthattheChinchillaRexrabbitpopulationwasatHardyWeinbergdisequilibrium,thewhiteRexrabbitpopulationwasatHardyWeinbergequilibrium.PICofchinchillaRexrabbitpopulationwasofmediumpolymorphism(0.25<PIC<0.50),whiteRexrabbitpopulationwasoflowpolymorphism(PIC<0.25).[Conclusion]TyrpgenehascertaineffectsonthehaircolorofRexrabbit.ThisresearchprovidescertainreferencesfortheselectionofhaircolorofRexrabbit.Keywords Rexrabbit;Tyrp1;SNPs;Sequenceanalysis

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Page 2: nop 1Tyrp1Eist £23 - nykxw.com · :2 uvÆ29 AA´ BBEi{p@smn Fig.2 ThesequencingresultofAAandBBgenotypesatexon2

4�·Q4

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1 x�`y

Table1 Theinformationofprimers

�;

Primer�;|�

Primersequence(5′→3′)�Ø%p

Fragmentsize∥bp

ì�#$

Annealingtemperature℃

1 F:TTTACAGCTGGACTTTGGTCTR:GCTTTGAACCTCATACAAGCCT

454 59

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323 58

3 F:ACAAGCAGAGAATACTAACAGAR:CTCCTAGCTGTAATTTGGAAC

205 57

4 F:TAAGATTCTAAGCGTTGACCTR:CTAGACCAGTTGGAAGCACAC

168 58

5 F:TTTGCAATAGTTTTTACTCTTCTR:CAGTGGCATCTTAATAATCCC

180 50

6 F:TAATATTAACAAGTTGCCTR:TAAAATACATTTCAATGCTT

147 48

7 F:TTTTCTTTTTCAAATAGGTCAR:CTGAATATTCGGACTCACA

206 51

1.2 VZT'

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1.3.2 +J

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Page 3: nop 1Tyrp1Eist £23 - nykxw.com · :2 uvÆ29 AA´ BBEi{p@smn Fig.2 ThesequencingresultofAAandBBgenotypesatexon2

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Table2 Biologicalinformationstatisticsofexon2ofTyrp1lociin2rabbitpopulations

(Q

Population7�ô

Samplenumber∥ô

@»×f&

Genotypefrequency

AA AB BB

_�@»f&

Allelefrequency

A BHe PIC χ2

C 392 0.215(84) 0.330(129) 0.455(178) 0.380 0.620 0.471 0.360 35.114W 174 0.779(127) 0.202(33) 0.018(3) 0.880 0.120 0.210 0.189 0.246 1

:C£TdUP.

,W£{}P.

;õ

df=2q

,χ2(0.01)=9.21,χ2(0.05)=5.99。

 Note:CwasChinchillaRexrabbits,WwasWhiteRexrabbit;whendf=2,χ2(0.01)=9.21,χ2(0.05)=5.99.

o@

,>ÃH�£�pD@»

。�

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0

(Arg+Lys)£

47,t�µòô>

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3 TYRP1pÒEÓ�Á

Table3 CompositionofTYRP1aminoacid

?F

Nameô0

Number"&

Ratio∥%?F

Nameô0

Number"&

Ratio∥%Ala(A) 34 6.1 Lys(K) 11 2.0Arg(R) 36 6.4 Met(M) 12 2.1Asn(N) 34 6.1 Phe(F) 30 5.4Asp(D) 29 5.2 Pro(P) 39 7.0Cys(C) 18 3.2 Ser(S) 41 7.3Gln(Q) 27 4.8 Thr(T) 37 6.6Glu(E) 27 4.8 Trp(W) 10 1.8Gly(G) 35 6.2 Tyr(Y) 19 3.4His(H) 18 3.2 Val(V) 31 5.5Ile(I) 24 4.3 Pyl(O) 0 0.0Leu(L) 48 8.6 Sec(U) 0 0.0

  vL

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1

:Ïáa}��@Óo@ô

,tØaJ}¶�D\Ï�

。0°)

£rÏ�

,0°'£\Ï�

Note:Horizontallinesindicatedtheaminoacidresidues,andtheverti

callinesrepresentedtherelativehydrophilicregions.Abovezero

wasahydrophobicregion,andthefollowingwasahydrophilic

area.

:

4 TYRP1ÒEÓst��k£;o

Fig.4 AnalysisofthehydrophilicpropertyofTYRP1amino

acidsequence

661           °±²³��

                         2016�

Page 4: nop 1Tyrp1Eist £23 - nykxw.com · :2 uvÆ29 AA´ BBEi{p@smn Fig.2 ThesequencingresultofAAandBBgenotypesatexon2

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