Upload
tranminh
View
216
Download
1
Embed Size (px)
Citation preview
nopÒÓ¡Ý6qr
1(Tyrp1)Eist;o�uvá.w£23
M N
1,2,ÛOP
1,2,QRÀ
1,2,#�S
1,2,Ä ?
1,2,þ T
1,2,UVW
3,X#¦
1,2 (1.?÷%&¸;h&#��
&'
,*�?÷
225009;2.*�.¸;òã#n�I;Ó·¿ª3
,*�?÷
225009;3.À*.¬@AA�.$Zµ:�
,À*¬@
315400)
45
[+,
]3YZ[\��`@]^
1(Tyrp1)�T�¦Ä_�F�û, <'i
。[D5
]ÔZ[
Tyrp1�TÊ`,a��Ê`
、b
c©�,�C|7�b¹PQd�V µ
,e2wx
PCRSSCP(Singlestrandconformationpolymorphismanalysisofpolymerasechainreactionproducts)
NÃË�
Tyrp1�T
CDSÊ`,<H� &f®
。[Z[
]#g� µvã
,TYRP1]^6åh_,i7�]^
,ñ#$
j
,kñ\��ù�l4m���Zno
。<H�Ë�vãp�
Tyrp1�T
CDS,
483bpãq�
1r
C→T,estú
,q-uß<
2c
,rÁ�,túvñw����,7ú
(AUC→AUT,xy��
)。HardyWeinbergÐøËJâÞ
,@z{ÄZ[õ|ã-}ÐøfH
,
^ÄZ[õ|ã-ÐøfH
。@z{[õ|]ûf<H#gmg
(0.25<PIC<0.50),̂ÄZ[õ|]~f<H#gmg
(PIC<025)。[
Zq
]Tyrp1�TÔZ[,¦Äñ0_,ÕÖ
。rstZ[ì]Z[¦Ä,LoLÝyz0_,ñò
。
678
Z[
;Tyrp1;SNPs;Ê` µ
9:;<=
S829.1 >?@AB
A >CD=
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
EFGH
ãj²³©³NÃ|A¼�W�
(CARS441);KL�æ�
���¼�@����+
(PAPD2014134)。IJKL
MN
(1992-),�
,���_�
,̂�st¦
,stDº
:R
�HIªÝo
。E¡¢£
,¤¥
,��
,��¦§�
,��
R�HIÝoª��st
。
MNOP
20160221
Tyrp1@»>B
1H3CÐ%D}¼@»
,KPW@»
DED�{#F�ÓH¢£
,»¤ºF£F�ÓH¶¡�{
1(Tyrosinaserelatedprotein1,TYRP1)[1]。×<&¦�n�
(}¼a;CÐDÇHËÌ
,+F�ÓH
、<G}¼ìÊH
U
5,6S?@HIJÓ\BH
,L´�(}¼D�Ð6Zá
nÓ¯D³�
,PX
Tyrp1@»DED
5,6S?@HIJÓ
\BH>F�ÓHÓ¯Dó¢»�
,#F�Ó
(Tyrosine,TYR)、
F�ÓH¶¡�{
2(Tyrosinaserelatedprotein2,TYRP2)
Ã��ÐF�ÓH¶¡�{-K
[2]。lZ56}
~
,Tyrp1@»ü�ØTI#(}¼DaÐcMè
,�MÛL
¾�µÞ*¶F�ÓHDûBbc
,I#VÀ(}¼pQD
�·
,ÞBCY(}¼@sDZTUÖ×
[3-4]。¥¦¹º
PCR��MZ
Tyrp1@»
7Hè`�D|�
,�
Tyrp1@»
�@Ó|�«¬�a;&?Nn�
,ÞÉ�
PCR-SSCP�
�UØT4|Dæ��6Hè`�«¬<¨c34
,n�
Tyrp1@»Ot¢O}P.
(Oryctolaguscuniculus)(QD<
¨c
,ßàP#P.¡}�ÐD¡b
,QO£P.¡}D®
`®ã�¡ÃµDIí
。
1 QRST'
1.1 VZQR
1.1.1 ©ª¸;
。XÞR�¢�na
、�äD
6ÃSTd
U}P.
392ô
、{}P.
174ô
,X
566ôP.³£©ª�
Í
,©ªHQOw�À*.¬@AA�.$Zµ:�
。
1.1.2 ©Ö
。�{H
K、10×TBE、Tris、EDTA、KHßà_
©ÖO��a;
()�
)7�Zµ:�
;10%ºVÓW
、
TEMED、SDS、y÷
、̀Î_©ÖO��"RX�a;��Z
µ¼½:�
。
1.1.3 �;I;
。89
NCBI,-):oD.
Tyrp1@»X
|�
(YÖ}
:NC_013669.1),n0O
7Hè`��8èD¡
U=���8I;ÞCоuc�QD�;
,�;Ka7a
;7�
()�
)Zµ:�CÐ
,�;?N�}
1。1.1.4 Tyrp1
@»|�n�êÂ
。É�
ExPASy7Y�@»
DED�{/c/«¬n�
,�
ProtParamn��{D�
@Ó|��Ð
、¶�n�/0U_ß·_ËBc/
,�,
)Oa7Y
SMART(http://smart.emblheidelberg.de/),n
�
TYRP1�{�·8
,Þg�
NPS@5Zå)D
SCOPMAn�Q4�{DS4�·
。É�
TMPRED(http//www.ch.embnet.org/software/TMPRED_form.html)
«¬�{/|�D
[� � n �
;& �
ProtScale(http://web.expasy.org/protscale/)
«¬�@Ó|�D\Ïcn�
。É�
SwissModel(http://swissmodel.expasy.org/)
�
TYRP1DE�{«¬Ø
abDc
þ��
abde
¶·[\]^_`
,JournalofAnhuiAgri.Sci.2016,44(9):164-167,196
4�·Q4
。
f
1 x�`y
Table1 Theinformationofprimers
�;
Primer�;|�
Primersequence(5′→3′)�Ø%p
Fragmentsize∥bp
ì�#$
Annealingtemperature℃
1 F:TTTACAGCTGGACTTTGGTCTR:GCTTTGAACCTCATACAAGCCT
454 59
2 F:TCCCACAAAGCAGATATCTCAR:CAGCAAAGGCATCTCGTCT
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'
1.2.1 DNA��
。����=
/¿]R����P.^�
r7@»�
DNA,g�
TE�wÀ
100ng/μL,g�
NanoDropND1000
ñÓ2$4µê4µ
DNA2$
,Ãè
ODGDwx
£
1.7~1.9,�Lá_D79M�
。
1.2.2 PCRMZ
。PCRcMQb
(20μL)£:10×PCRBuffer2μL、10mmol/LdNTPs2μL、10pmol/L)、'��;/
1μL、5U/μLTaqH 0.2μL、100ng/μLDNA�`
1μL、ddH2O128μL。PCRcMÁÂ�'
:95℃QÊc
5min;95℃
Êc
30s,�`D#$ì�
30s,72℃=a
30s,35H/
º
;72℃±Z=a
10min,10℃'�N
。
1.2.3 PCRSSCP34
。O
20μLPCRMZ¶;X1Æ
5μL)7o²!
,�CQbO
98℃Êc
10min>
,�P
-20℃'}Xðï
5min,P
10%ÚÊctKHßàbMß
£cÙ·7
,±�
250Vߣ
,10min>òߨ*Ù
120V,ߣ
12h。d<`}>*ú>ïZt¢²×
。IZt¢²
×n\
,òP
PCR¶;eÙa7a;7�
()�
)Zµ:�
4|
,°Þµ{@ÏÊDA×
。
1.3 Zk;o
1.3.1 @»f&U@»×f&
。@»f&
(Genefrequency)>ò(QX;þµ_�@»#Py_�@»D"&
;@»
×f&
(Genotypicfrequency)>ò(QX;¾µ@»×HQ
ôgX HQôD"&
。
1.3.2 +J
-#h
(HardyWeinberg)áG3ª
。
χ2=(O-E)2
E (1)
ìX
,O(Observed)J}6H@»×D*4ôõ
;E(Expected)
J}6H@»×O+J
-#háGµiÐ�Djµ'
D[âôõ
。
1.3.3 �C$
(Heterozygosity,He)。�C$+@»<7
c
[3],[â�C$D;<:ì£
:
He=1-k
i=1p2i (2)
ìX
,He£[â�C$
;k£_�@»ô
;pi£B
kHk�B
iH_�@»Df&
。
1.3.4 <¨?N�0
(Polymrphisminformationcontent,PIC)。
�7°':ì;<<¨?N�0
(PIC):
PIC=1-k
i=1p2i-
k-1
i=1
k
j=i+12p2ip
2j=2
k-1
i=1
k
j=i+1pipj(1-pipj)(3)
ìX
,k£_�@»ôõ
,piUpjn0£B
iUB
jH_��
ØDf&
。
2 mnS;o
2.1 Tyrp1EiXuvápz°mn
�W@»D
7Hè
`�|�«¬MZ
,Þ�
2%D®�øbMߣ34
。34
MZ�Ø%pr�D
MarkerO£
DL500Marker。vL
1M
°xn
,Tyrp1@»/è`�DÁ²¾ucO�Q
,�²�
¢
,¾uÁ²D�Ø%p#ËÌGÃþ
。�
PCRSSCP34
>
,O
Tyrp1@»è`�
1、3、4、5、6、7XO>34À<¨�
·
,íOè`�
2)34À
2H_�@»
、3̀@»×
(AA、BB、AB)。
:
1 Tyrp1Eiuvá
2̂¾Ei{påæ:Ú
Fig.1 TheSSCPelectrophoresisimageofdifferentgenotypesof
Tyrp1exon2
2.2 Tyrp1Ei.wst;o
g�
AlignIRV2.0êÂ
,ò
4|¿ÀD
DNA|�«¬�"
。vL
2~3M°xn
,Tyrp1@»è`�
2O
29bp(·H@»
CDSD
483bp)ñZ
1H
C→TDÏÊ
,{@DÏÊÞÓZ,Ð�@ÓD�Ê
(AUC→AUT,
ul�Ó
),£¢ªÏÊ
。
2.3 Tyrp1Eiuvá
2p|}.y£;o
{}UTd
U}P.è`�
2/@»×U@»×f&�mæ�Cc3
ª
,BB@»×f&OTdU}P.X�Ø
,O{}P.X�
í
,}~_�@»
B>TdU}P.(QXDJ__�@
»
,dO{}P.X_�@»
A>J__�@»
。HardyWeinberg
áG3ª}~
,TdU}P.(QñPÚáGi
¨
,{}P.(QñPáGi¨
。TdU.(Q£X$<¨
?N�0
(0.25<PIC<0.50),{}P.(Q£í$<¨?
N�0
(PIC<0.25)。TdU}P.�C$�Ø
,{}P.
�C$�í
(}
2)。2.4 Tyrp1
Eist;o
2.4.1 TYRP1�@Ó|�DËBc/
。89
GenBankô9
n)«È}D-.D
Tyrp1@»|�MN
,-.D
Tyrp1@
»�P
1}<}Q)
,Xz
15843bp,Z
7Hè`�U
6H
=��
。DE|�K
1683H{@�Ð
,DE
560H�@Ó
56144·
9̧ M N¹
Z[\��ù`@]^
1(Tyrp1)�TÊ` µ�uß<<H�st
:
2 uvá
29
AAÂ
BBEi{p@smn
Fig.2 ThesequencingresultofAAandBBgenotypesatexon2
:
3 ^¾Ei{Iuvá
2~�Óstp��
Fig.3 Homologouscomparisonofnucleotidesequenceofexon2indifferentgenotypes
f
2 2��o��
Tyrp1uvá
2x.w��p��``y��
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@»
。�
ProtParamOaêÂQ4
,
n�ì£
C2802H4268N786O831S30,ï�Eô£
8717H
,n�0
£
63223.2kD,ËÌ_ß·£
6.06。OrZDE�@ÓX
,
Leu(L)�0±Ø
,£
8.6%,Trp(W)ôg
1.8%(}
3)。²W
ßqo@DEô0
(ASP+Glu)£
56,²½ßqo@DEô
0
(Arg+Lys)£
47,t�µòô>
52.92,�~XH�{/t
�µ
。
f
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
4M°xn
,TYRP1�{rÏc±%G£
3.711,±
pG£
-3.133。TYRP1Dy¯\ÏáOô
(GRAVY)G>
-0.381,KP
GRAVYGDwx·P
-2#
2ml
,P£½
Gq}~¤�{£rÏ�{
,dWG}~£\Ï�{
,»¤
W�{>\�{
。¢q
,TYRP1�@Ó|�=s%<ô£
\Ïco@
,}~
Tyrp1@»DE�{£Ïÿc�{
。
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�
2.4.2 TYRP1�@Ó|�DS4�·UÇ4�·Q4
。¹
º
NPS@5Zå)D
SOPMA�
TYRP1«¬S4�·Q4
,
�V�}
4r�
。PX
,TYRP1�{S4�·°αuâUÎ
¡¥vÊ£y
,ZÉP�µ�{/D�·
,Ρ¥vÊ>
TYRP1�{±%D�·iÂ
。X}~
TYRP1�{DS4�
·£�C×
。
vL
5M°xn
,Ç4�·|�D¶G$uÀ
84.17%,Evalue
G£
0,f~�×#�¾DQ4�{|�w³c
�Q
。
f
4 TYRP1qrpô�m�;o
Table4 AnalysisofTYRP1secondaryproteinstructure
?F
Name
SOPMAô0
Number"&
Ratio∥%αuâ
Alphahelix(Hh) 191 34.11310uâ
310helix(Gg) 0 0Pi
uâ
Pihelix(Ii) 0 0β�|
Betabridge(Bb) 0 0=ax
Extendedstrand(Ee) 106 18.93βãÛ
Betaturn(Tt) 41 7.32yÊ
Bendregion(Ss) 0 0Ρ¥vÊ
Randomcoil(Cc) 222 39.64tԵ�
Ambigousstates 0 0Py
Otherstates 0 0
:
5 TYRP1p��m�·{
Fig.5 ThetertiarystructuremodelofTYRP1
2.4.3 TYRP1�@Ó|�D%Ûc�·8Q4
。É�
SMART(SimpleModularArchitectureResearchTool,�I�¯
·z{à7Y
)n��V`�
TYRP1�{Z
3H%Ûc�
Ø
(L
6)。d}�
(1~47)J}?}|
;Ù}�
(205~440)J}F�ÓH-K¾¿c�8
;U}�
(502~524)J}[
��
。
3 K�Sm�
%056}~
,Tyrp1@»#¸;D¡ÈO}U}}¶
¡
,W@»DED
5,6S?@HIJÓ\BH
,>F�ÓH
ӯD�
,O(}¼D�ÐXYZánÓ¯D³�
。
:
6 no
TYRP1qrstp
SMART;omn
Fig.6 SMARTanalysisresultsofTYRP1inrabbit
�A
Tyrp1@»è`�
6)D
1H{@Jk
,þgÇA~�
���
,ýþ
TYRP1�{ékbc
,g¿P�F�Ó?BH
DûB³�p�
,���S�{BÌ
3(OCA3)[5]。Rieder_
[6]56È\Ù}RS�X
Tyrp1@»D}u0�ÚÙ}R
í
,@4
Tyrp1@»#RD�cÙ¡}ci¶¡
。Li_
[7]5
6È\
,�t¢}}X
Tyrp1@»D}uNO�u
,(}�X
Tyrp1@»D}u0£{}�D
100008°)
。56}~
,
Tyrp1@»DED
TYRP1<G}¼ìÊu·H
(DCT)�
ûB<G�ãB£1(}¼
,BC(}¼pQDÐU(}¼
@sDZT#Ö×
,lMØTI#(}¼DaÐ
[8]。
£�ô�
Tyrp1@»O¡}�к�XD³�ÞÑ
,¥
¦���P.
Tyrp1@»|�
,�
Tyrp1�@Ó|�DËB
c/«¬n�
,ÜÄ�µW�{£\Ït�µ�{
,1)S
4�·��Qn�
,M°�W�{D�·�%Û«¬ÜÄ@
4
。¥¦�
TYRP1�{�@Ó|�Dn��V#«ZDÖ
¸
[9]¶�C
,W�{£
I×[�ø�{
,µ�O(}¼QD
��)
,¶õ% n�8ÿP(}¼Q�=DÿM=
,I8
7³t�µ
,ͯ¹ºA}Saz»�@|_¢Py�{�
F�ÓH�CDæì�µ7³
,αuâUΡ¥vÊ>
TYRP1�{|�±%0D�·iÂ
,�oP·H�{/X
;
SMARTn�`��W�{D
3H%Ûc�Ø
,+
NpD?
}|
,Xl¶õ%�ØDF�ÓH¶¡-K¾¿c�·8�
CpD[��·8
。
õ�
,×<�=è&¦56
Tyrp1@»D<¨cO¡}
�к�XD³�
。+�_
[10]56È\��R
Tyrp1@»
è`�
2D
1H�ªÏÊ#Å}¡}¶¡
。Rieder_
[11]5
6È\R
Tyrp1@»è`�
2D
SNP(C189T)����ÓÀ
��ÓD�ªÏÊ
。Gratten_
[12]�(}U�}�!
Tyrp1@»D56È\
,è`�
4D
869bpñNO�ªÏÊ
(T869G),�}à�!@»×X£
TT,(}à�!@»×£
GGð
GT,}~W�·DÏÊ#à��!¡}¶¡
。�
�
[13]�
3̀}}�õ��
Tyrp1@»
PCRSSCPn�È\O
Tyrp1@»NO
2H<¨c�·
(T367CU
C1153T), >>
È\X
2H<¨c�·#�õ��}}mlNO`6¶¡
c
。W56X�
Tyrp1D
CDS|�D<¨cnoiÉ«¬
34
,34�P.
Tyrp1@»
7Hè`�D<¨c
,OP@»
è`�
1、3、4、5、6、7X>34À<¨�·
,íOè`�
2D
29bp(·H@»
CDSD
483bp)ñZ
1H
C→TD¢ªÏÊ
。
½�<7cn�È\
,TdUP.(QD
HeGØP{}P
.(Q
,f~L´YZV£��D½�<7c
,VQD®.
ÝE
。<¨?N�0
(PIC)>�wG0@»�Ø<¨cDÓ
(èLµ
196¶
)
76144·
9̧ M N¹
Z[\��ù`@]^
1(Tyrp1)�TÊ` µ�uß<<H�st
3 Î]à^_T`bßÎúpeq
3.1 ¤r�`bsVté
ØÀg�´v���
、íf&
D*F�ãFU��
、Øf&D ë�
,,ã�ÃH@ô�
%RÈÒcÙD&Y�AB
。C-ØÀòÇ.«�6XÆ
�
“4hY��
”,��D
“za�
”lëìF
“�a�
”、“ú(
2~3��
”_&Y��ìãÊ
,Y�f&ëìZ1
,�PY
�õD|D��ÙÚE0l[\n¶õ%D{zAM
。
3.2 u)¿4l&úplvp¨�
?NqJ
4G{Û×
ÞDY�gYc<¹º,p
、×Þ
APP_ß�mZáë�¶
Y�¶9
。ØÀ-
、«�l¯1ÿØÀY�¨¸{ÛD?N
,p
。*.`�
,Yc�P&?Dg�&�Ø
,»¤MO:
W3
、�Wk?¤Õ_I��a/Y�õD|Y�à�DS
VE
,�Yc¹º��SVE¡1/Y�õD|:Xáë
,
I#&?ì¸
,vduÀÅ$��DAV
。zs1ÿ#/Y
�,-DC³
,O¨¸{Ûáë.I)
,Å$XC&?
、&©
_=¾êÂ
,�qn|/«�Y��õ
、Y�yýb¸_
,@
¸Y�hÌæìDã×]4
。
3.3 wxà^¼yæ�¬æ
ØÀ£�aY�õD|²w
Ø9/DcD
。89Yc¾·
,MvY�IÕUY�5Zâ
Hæ竬t�
,�M\JØÀY�ABDͯ
,ǹ/Ø
À-Ù«�DY�©a
,�¡ØÀY�
“عW
”,¿\Yc
“���
”。
3.4 ¬zÖ{`bÎI²Ö
1ÿY�C³
,�ÐY�X
¢Q
。��
、Y�y¢ 5¯1ÿ¾D
,�.�aY�¹�
,
�Y�ࣾ}
,�ÐY�à£ìýUY�ÈÒCE
,¿
\�8Y�Dã×]4
,�E¿\|�Y�ÃQB
。.�Y
�à£UY�5ZX|ÞÑ
,¿\/«�ÎÒ�T
,1½ò
“¢�AM
”&�ÀY�¿ËX
。
4 m�
¥¦@PC-ØÀ�c*.ô9
,56�C-ØÀǹ
>��cY�¦)¬£DBC
, µP7�ô0Zµ
,�Ì
DYZ��cUM�@LcZØZÿ
,�ØÀ#�cY�¬
£D=OBCÞÑJKÅ|z56
,XO>Á*.XZØ«
ÃÄ56
。
��>?
[1]KOBAYASHIK,OKXIMURAM.Thegrowthofcitysystemswithhigh-speedrailwaysystems[J].Theannalsofregionalscience,1997,31(1):39-56.
[2]ÂÝø
,�c
,�;º
,Z
.ÉÙÞßàÞá923
:à�OVÉÙâÞ
o�tJî
[J].̂:ã¬
,2015(7):8-12.[3]
�ïû
.ÙäàÞå0æçRQh©Æ
[J].àÞ1F
,2015,30(1):8-10.
[4]��
,èÆ7
,®VV
.$wÉÙéßá9ê�RàÞ�5Jî
[J].*
ë23R��
,2014,33(2):94-98.[5]
�5Ô
,�¬¾
,�ãº
.ÉìÙä�o�í&#Þá9tL[23
:
8³"�9F
[J].*ë23R��
,2014,33(4):121-125.[6]
èÆ7
,��
,Õl�
,Z
.ÉÙéßtàÞ~�RwQê�Jî
[J].!"îýï�ð1ê1y
,2013(11):63-71.[7]
®lÆ
,���
,O��
,Z
.o}ÉìÙä�o�àÞß.tL[
:8
³"�9F
[J].BC*Â
,2013,33(7):163-168.[8]
>,¾
,Ó~
,ØP
,Z
.JëàÞ.ѨöàtÉÙä\-CÂ
:8Ò
�"ñÉÙ9F
[J].*Â1y
,2015,70(2):214-233.[9]
>,¾
,Ó~
,�Ì
,Z
.�ïÉìÙä�àÞL[t23-òó
[J].*Âi1
,2012,32(3):
檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪
322-328.
(c´µ
167¶
)
¯òù
,Oè`�
2D@»×X
,TdUP.(QD
PICG
·P
0.25#
0.50ml
,è`�
2@»k[X$<¨
,f~W
(Q�Pè`�
2@»×D®.YZõDÝE
,X�V
#@»�C$D56�V@�Ãþ
。¤è
,TdU}P.(
Q��+J
-#ÌáG
,M°ÜÄ�µW�·DÏÊZÉ
P1(}¼DCÐ
。@»�@»×O(QXDno¶Él
DÀW@Ì
,»¤
Tyrp1@»�P.D¡}ZõBC
。
��>?
[1]�ô
,Êõ�
,®tö
,Z
.TYRP1à:ͶÜ(LÔ,It2345
[J].Ò�Þß÷2
,2009,36(9):94-96.[2]
�
,!5ø
,��è
,Z
.ù45vK�×¹
1ÌÍíLÔ,It23
45
[J].ÞßRà�i1
,2010,31(10):114-116.[3]KOBAYASHIT,HEARINGVJ.DirectinteractionoftyrosinasewithTyrp1toformheterodimericcomplexesinvivo[J].JCellSci,2007,120(Pt24):4261-4268.
[4]RADHH,YAMASHITAT,JINHY,etal.Tyrosinaserelatedproteinssuppresstyrosinasemediatedcelldeathofmelanocytesandmelanomacells[J].ExpCellRes,2004,298(2):317-328.
[5]BOISSYRE,ZHAOH,OETTINGWS,etal.Mutationinandlackofexpressionoftyrosinaserelatedprotein1(TRP1)inmelanocytesfromanindividualwithbrownoculocutaneousalbinism:Anewsubtypeofalbinism
classifiedas"OCA3"[J].AmJHumGenet,1996,58(6):1145-1156.[6]RIEDERS,STRICKERCH,JOERGH,etal.Acomparativegeneticapproachfortheinvestigationofageinggreyhorsemelanoma[J].JAnimBreedGenet,2000,117(2):73-82.
[7]LISJ,WANGC,YUW,etal.IdentificationofgenesrelatedtowhiteandblackplumageformationbyRNASeqfromwhiteandblackfeatherbulbsinducks[J].PloSone,2012,7(5):36592.
[8]AROCAP,URABEK,KOBAYASHIT,etal.Melaninbiosynthesispatternsfollowinghormonalstimulation[J].JBiolChem,1993,268(34):25650-25655.
[9]SARANGARAJANR,BOISSYRE.Tyrp1andoculocutaneousalbinismtype3[J].PigmentCellRes,2001,14(6):437-444.
[10]LIB,HEX,ZHAOY,etal.Tyrosinaserelatedprotein1(TYRP1)genepolymorphismandskindifferentialexpressionrelatedtocoatcolorinMongolianhorse[J].Livestockscience,2014,167:58-64.
[11]RIEDERS,TAOURITS,MARIATD,etal.Mutationsintheagouti(ASIP),theextension(MC1R),andthebrown(TYRP1)lociandtheirassociationtocoatcolorphenotypesinhorses(Equuscaballus)[J].MammGenome,2001,12(6):450-455.
[12]GRATTENJ,BERALDID,LOWDERBV,etal.CompellingevidencethatasinglenucleotidesubstitutioninTYRP1isresponsibleforcoatcolourpolymorphisminafreelivingpopulationofSoaysheep[J].PRoySocLondBBio,2007,274(1610):619-626.
[13]_ú
.TYR、TYRP1à:R�gûüýLK�H23
[D].þû
:7³
ijx1
,2014:29-39.
691 °±²³��
2016�