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2011,24(4):423 427
ForestResearch
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:10011498(2011)04042305
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:20110129
EFGH
:
(+n¡EF
üÚEF
(31000983);
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(2010ZC235)
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RTPCR
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:
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Rô67
JF731244。
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cDNACloningandSequenceAnalysisofβ1TubulinfromChinese
WhiteWaxScaleEriceruspelaChavannes(Homopetera:Coccidae)
YANGPu1,CHENXiaoming1,ZHUJiaying2,DINGWeifeng1,LIUWeiwei1
(1.ResearchInstituteofResourceInsects,ChineseAcademyofForestry,TheKeylaboratoryofCultivatingandUtilizationofResourceInsectsof
StateForestryAdministration,Kunming 650224,Yunnan,China;2.KeyLaboratoryofForestDisasterWarningand
ControlofYunnanProvince,FacultyofConservationBiology,SouthwestForestryUniversity,Kunming 650224,Yunnan,China)
Abstract:AfullengthcDNAofβ1tubulingenefromEriceruspelawasobtainedbyusingRACEandRTPCR
methods.Thesizeofβ1tubulingeneis1492bp,whichcontainsanopenreadingframeof1344bpthatencoding
447aminoacidresidueswith5020kdamolecularweightand475isoelectricpoint.Theputativeaminoacidse
quencescontainconservedsequencesofβ1tubulinandtheGTPbindingsite.Thephylogeneticanalysisshowedthat
β1tubulingeneofE.pelashareshighidentitywithβtubulinhomologuesfromotherinsects.Thesequencesofβ1
tubulingeneofE.pelaweresubmitedtoGenBankandtheaccessionnumberwasassignedJF731244.
Keywords:Ericeruspela;tubulin;RACE;aminoacid
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。
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J
5’RTPCR
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wD.Rô β1)¿ ESTÐ:EÖv
f
,
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zÜÉ
3’GSP(5’TGTCTGGAGTCAC
TACCTGTCTACG3’)
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zÜ@_
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PCRMasterMix(
ðw
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3μL,cDNA3μL,ddH2O|¢ 50μL。
PCR
nÈÏÐ
94℃ 4min;94℃ 30s;50℃ 30
s;72℃ 90s,31
|}
;72℃
~
10min。
wD
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zÜ{f
,
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AGTGATGAAGAAAC3’)
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f
,
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1
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cDNA
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。
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tp://www.ncbi.nlm.nih.gov/)
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U
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Ò
1)。
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NÌ;K
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0E1Ð:
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96% 998%,
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ã
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>
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zÜ
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r9
2
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C8
:
2000、1000、750、500、250、100bp。
1 AAAATGAGGGAAATCGTACATCTGCAAGCCGGTCAATGCGGTAATCAAATCGGAGCTAAGTTTTGGGAAATCATCTCGGACGAGCACGGC
1 M R E I V H L Q A G Q C G N Q I G A K F W E I I S D E H G
91 ATTGACGCTACTGGAACCTATCATGGCGACTCCGATTTGCAGCTGGAAAGAATAAATGTGTACTACAATGAAGCTTCAGGAGGTAAATAT
30 I D A T G T Y H G D S D L Q L E R I N V Y Y N E A S G G K Y
181 GTTCCACGTGCCATACTGGTCGATCTGGAACCAGGTACTATGGATTCAGTTCGCGCTGGACCATTCGGTCAGATATTCCGCCCTGATAAT
60 V P R A I L V D L E P G T M D S V R A G P F G Q I F R P D N
271 TTTGTATTCGGACAATCGGGAGCCGGTAACAATTGGGCCAAAGGACATTACACCGAGGGTGCCGAGTTAGTCGACTCCGTTTTGGATGTT
90 F V F G Q S G A G N N W A K G H Y T E G A E L V D S V L D V
361 GTTAGGAAAGAAGCCGAAAGTTGTGATTGCTTGCAAGGTTTCCAACTTACACATTCTTTGGGTGGTGGTACCGGATCCGGTATGGGCACC
120 V R K E A E S C D C L Q G F Q L T H S L G G G T G S G M G T
451 TTGCTTATTTCAAAGATCAGAGAAGAATACCCCGACAGAATCATGAACACGTACTCCGTTGTACCTTCGCCAAAAGTATCCGACACAGTA
150 L L I S K I R E E Y P D R I M N T Y S V V P S P K V S D T V
541 GTTGAACCGTACAATGCCACGTTATCAGTTCACCAACTCGTTGAAAACACCGATGAAACGTACTGCATAGATAACGAAGCCCTATACGAC
180 V E P Y N A T L S V H Q L V E N T D E T Y C I D N E A L Y D
631 ATTTGTTTCAGAACGTTGAAACTTACGACGCCCACCTACGGTGACTTGAACCATCTCGTATCCCTGACCATGTCTGGAGTCACTACCTGT
210 I C F R T L K L T T P T Y G D L N H L V S L T M S G V T T C
721 CTACGATTCCCCGGCCAACTGAACGCCGATTTGCGTAAACTGGCCGTCAACATGGTTCCCTTCCCACGTTTACACTTCTTCATGCCGGGC
241 L R F P G Q L N A D L R K L A V N M V P F P R L H F F M P G
811 TTCGCTCCACTCACGTCGAGAGGTAGTCAACAGTACAGAGCACTCACCGTTCCTGAATTAACGCAACAGATGTTCGATGCCAAGAACATG
270 F A P L T S R G S Q Q Y R A L T V P E L T Q Q M F D A K N M
901 ATGGCCGCCTGTGATCCCAGACACGGAAGGTACCTAACCGTCGCCGCCGTATTCCGAGGACGTATGTCGATGAAGGAAGTCGACGAACAA
300 M A A C D P R H G R Y L T V A A V F R G R M S M K E V D E Q
991 ATGTTGAACATCCAAAACAAGAACAGCAGCTACTTCGTCGAGTGGATCCCCAACAACGTTAAGACCGCCGTTTGTGATATTCCACCACGA
330 M L N I Q N K N S S Y F V E W I P N N V K T A V C D I P P R
1081 GGATTGAAGATGTCAGCCACTTTCATCGGCAACTCGACCGCCATCCAAGAATTATTCAAACGTATCTCCGAGCAGTTCACCGCCATGTTC
360 G L K M S A T F I G N S T A I Q E L F K R I S E Q F T A M F
1171 AGAAGAAAAGCTTTCTTACATTGGTACACCGGTGAGGGTATGGATGAGATGGAATTCACTGAAGCCGAATCCAACATGAACGATCTGGTA
390 R R K A F L H W Y T G E G M D E M E F T E A E S N M N D L V
1261 TCCGAATACCAACAATACCAAGAAGCTACTGCCGACGAAGATGCCGAATTCGAAGACGAACAAGAACAAGAGGTCGATGAAAACTAATAT
420 S E Y Q Q Y Q E A T A D E D A E F E D E Q E Q E V D E N
1351 TCCACTTTAAACGTAACGCTTATTTAATATTTTTTCCAGCCTGAATTTTTCCCTTTTTATTTTTTTTTTTTTATATGTATTTGTATTTAG
1441 AAAATAGAAACTGATTTACCATATAAAAAAAAAAAAAAAAAAAAAAAAAAAA
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,
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2
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$
%
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24
ü
cF
NCBI
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:Bombyxmori[AB011069],Mythimnaseparata[EU234504],Manducasexta
[AF030547],Laodelphaxstriatelus[AY479977],Monochamusalternatus[EU373305],Aedesaegypti[DQ833528],
Glosinamorsitans[DQ377071],Drosophilamelanogaster[NM079071],Maconelicoccushirsutus[EF070480],Culexpipi
ens[DQ401465]
Ò
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,
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,
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C
¬
[2,14-15]。
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MREI
]·Ð:UEÖ
5’
f
,¨
©
5’
RACE
ª«¥Ï
,
ô?Rô¿&õ β1)¿
5’
Ð:
。
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3’
WA¬EÐ:
,
0®gf./¤{A
Ð:
,
¡Ý%&oé÷ø^E03WA
447
|0E1B¯
。
Ð:
,
m%&Rô
Ð:ñβ)¿E0Ð:。
Ì;K
:
¿&õ β1)¿}Ã
ôõ0E1Ð:4sK/¼
,
0M1Ð:¬¦=
,
®ÿéÌcβ)¿FèÅØ]·K,
1ut°M}?u]·K
。
m%&z¿&õ
}Ãβ)¿0E1Ð::¿&õ-
¡¢q£T¤5_(Ò
,
\$¿&õ-
¡¢q£T¤5_¦Ò
,
}à±òP®²±*
+¿&õ}Ã)¿E0
cDNA
Ð:
,
ÊÉ
)¿]·Kzôõ~eôõ}ÃE
è5_L
。
m%&*+¿&õ
1
|)¿E0
,
)%
ý(pó;ôõû,|T*+ β)¿,
ÝFF¥³´
(Drosophilamelanogaster)
.NØ
4
cα)¿5c β)¿E0T*+q
U
,
ÃPã½ôõéÌ~/µLéÌ
H
[15-17];
F+¶
(Bombyxmori)
Ø
3
c α)
¿
4
cβ)¿E0T·qU[18],}1c
β)¿E0%Fè¸D¹ºvã
[19],1
cβ)¿E0»¼ôØ5,}
ãiabw q
[20],1
c α)¿E0
»¼~/7ØÞ.45
[21];
F½¼
¾
(Heliothisvirescens)
½¿ð¾
(Manducasexta)
)*+q
1
c β)¿E0,}½¿ð
¾-À·qβ)¿E0%1cv
ã β)¿E0,~ÑÃôõabØ
5
[22-23]。
0w
,
z½¿&õ@Ø}Ã)¿e
}?uðÁ\L=]%&
。
ef!A
:
[1]
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L
,
ÃaÄ
,
ÅÆÇ
,
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.
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Ð:*+Ð:
[J].
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,2008,34(4):130
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[2]
½ÉÔ
,
Ï
,
À
#
,
ä
.
Ê\<ËÌÍØ β1)¿E
0
cDNA
W²*+e}]ã
[J].
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,
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724