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Cloning and Expression Analysis of Actin ( MwACT 2) in Agropyron mongolicum Keng

蒙古冰草肌动蛋白基因(MwACT2)克隆与表达分析



全 文 :!24" !1#
 Vol.24  No.1
! " # $
ACTA AGRESTIA SINICA
   2016$  1%
  Jan.  2016
犱狅犻:10.11733/j.issn.10070435.2016.01.018
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(犕狑犃犆犜2)ÌÍW`Ô>r
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78

»/0ŠdRTPCR6RACE¾¨,Í34|-(犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng)&=5d1M9!ó¦®
5

ýïn犕狑犃犆犜2。lj=•gh:犕狑犃犆犜2®5àŸ1506bp,»J1M1131bpT¼>F(Éù,犕狑犃犆犜2
ó¦ŸAn376M…®WI®,=m`n41.606KDa,ùy?`­n5.29,»JactinÐ=Ÿ­,¬éactin¬’
¯

Y®5³)T…®WljGenebank&Ýa¢ë(犜狉犻狋犻犮狌犿_狌狉犪狉狋狌)9!ó¦(ÙN{:EMS62134.1)…®W
ÞJ99%Tß#,¬éȸó¦。Ïdˆ©`RTPCR¾¨=• 犕狑犃犆犜2®5TgW,efghY®5g
W¨©

Às)

š‰

€ÛÓù_TgW–Aغ

9:;

34|-

9!ó¦®5

®5d
;5'RACE;9KÔ:+=•;gW=•
<=>?@
:Q71    ABCDE:A     AFG@:10070435(2016)01012908
犆犾狅狀犻狀犵犪狀犱犈狓狆狉犲狊狊犻狅狀犃狀犪犾狔狊犻狊狅犳犃犮狋犻狀(犕狑犃犆犜2)犻狀
犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 犓犲狀犵
AOTegenbaiyin1,5,GAOLijie5,ZHANGShuhua2,YUNJingfeng3,LANGMinglin2,YANGXueju2,4
(1.ColegeofAgronomy,AgriculturalUniversityofHebei,Baoding,HebeiProvince071000,China;
2.ColegeofLifeSciences,AgriculturalUniversityofHebei,Baoding,HebeiProvince071000,China;
3.ColegeofEcologyandEnvironment,InnerMongoliaAgriculturalUniversity,Huhhot,InnerMongolia010019,China;
4.KeyLaboratoryofCropGermplasminHebeiProvince,Baoding,HebeiProvince071000,China;
5.ColegeofAnimalScienceandTechnology,AgricultureUniversityofHebei,Baoding,HebeiProvince071000,China)
犃犫狊狋狉犪犮狋:AnActingeneof犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 KengwasisolatedbyRTPCRandRACE,wasnamed
as犕狑犃犆犜2.Sequenceanalysisindicatedthatthefullengthof犕狑犃犆犜2geneis1506bpandcontained
an1131bpopenreadingframe.Itencodesapolypeptideof376aminoacids.Theestimatedmolecular
weightoftheputative犕狑犃犆犜2proteinwas41.606kDawithanisoelectricpointof5.29.犕狑犃犆犜2hasa
highlyconservedactindomain,whichdemonstrateditisamemberofactinfamily.犕狑犃犆犜2waspredic
tedtobeanontransmembraneprotein.Thephylogeneticrelationshipalsoindicatedthat犕狑犃犆犜2protein
wassimilarto犜狉犻狋犻犮狌犿狌狉犪狉狋狌(accessionno.EMS62134.1)withaminoacidsequencessimilarityof99%.
ResultofsemiquantitativeRTPCRshowedthattheexpressionof犕狑犃犆犜2wasconstant.Therewasno
significantdifferenceunderhighsalt,droughtandcoldtreatment.
犓犲狔狑狅狉犱狊:犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng;Actingene;Genecloning;5'RACE;Bioinformaticsanalysis;
Expressionanalysis
HIJK
:20141223;LMJK:20151017
NOPQ

z:ž*¾¿À´µ
(06220114D);z:žPo()¤)¾¨N]²´µ(1004002);z:ž«ïHµÁÂɒ¿£²³
(0901083);z:9Šd®¯/0²³•­®¯´µ(14966318D)“Å
5RST

À8}¦§
(1975),—,34,234J¶É,ÊË,²Y,ÍÎ?-¼°JG/0,Email:atg12@sina.com;<Ôc™ Au
thorforcorrespondence,Email:shmyxj@hebau.edu.cn;langminglin@tsinghua.org.cn
  9!ó¦(actin)wx+L门9K&,Úp
’H.DúTÌ«’=

Ls?JKN2
,actinÌ
«žH.ep

H.2û!6H.=R?9ùÉ
!
[1]。
jactin6?øó¦n®¯,6@ÄA_UV
ìH.ìT9Kš’

t9PR

Ô{+"

.E.
Fcdj|TGHÏd?
[25]。
P£JKTactin
Tĉó¦

ÞJsATÐ=6gW¨©

ÜG
9K%actin…®WljT@”sW70%~
100%[67]。
! " # $ !24"
èéactinLÜGä†&ΩgW,P/cnß
G•«TT¬®5dj¸t6@ç”TµT®5g
W`Tغ

ÚµvJK®5gW=•&PdT2
žãß

/0gh
,actinÚ荮5¬³)
T
[89]。
¶·n[

þÀÍps?JK

³´Q§
(犗狉狔狕犪狊犪狋犻狏犪)、»z(犣犲犪犿犪狔狊)、x9(犃狉犪犮犺犻狊
犺狔狆狅犵犪犲犪)、G Í N H I (犆犪狉犪犵犪狀犪犽狅狉狊犺犻狀狊犽犻
犻犓狅犿)、*0(犛狌犪犲犱犪犺犲狋犲狉狅狆狋犲狉犪犓犻狋狅犵)、?¸-
(犕犻犮狉狅狌犾犪狊犻犽犽犻犿犲狀狊犻狊犎犲犿狊犾)、eJ-(犣狅狔狊犻犪犼犪
狆狅狀犻犮犪)?&dd¡actin®5[1016]。
34|-
(犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng)ÞJ
ûsT‹dM

ÚÜà¬ê1ø&TH??-

@
2]Jû`Tø‰

ŽÐ+

Žx”T8

Ldé
ÜÐê+

УQÙ

cnß}cK

~u^

Y^
?

T¾9ä
‘ÞJGLdécK;ÒTø
®5

~ø‰

ø¥

Ž)?
)[1719]。
Ïd=mj|
®5acØ,;
—kóùKT
GF“”þ’n?-/0Tv’

»BCLž^þ°gT34|-9!ó¦®5
´,
[1819]
T®¯S

<=RACE¾¨—k34|-
9!ó¦®5àŸcDNAlj,\ßëì¡?-
9!ó¦/0T“ý˜

n?-=m9K+/0›
œ¡®¯‡ý

1 UVWXY
1.1 UVWjÏ
34|-Gmè234()Y+?-BCқ
œ

Y0®Þ0cTransT1、d/NnPEASY
T1,PCRMix1¶à⸺p,c RNA›fBÜ
ß
、TIANScriptcDNA!ßàš’BÜß1¶á}
9Kºp
,SMARTRACEcDNAAmplificationKit
(Clontech)1¶ TaKaRa,IPTG、Xgal?PdT
BÜ1¶Sª9Ã9KÃľ¨¼JAºp

Æ
L.ºpœ’ÊKš’6ÃlÃc

1.2 UV2ȇ
dzQÐ34|-Gm¼%

ºG%

bGé
»-èTéL&

Âé‘Ò“Dà

“DÍÖn
28℃,16h‘Ò/8h›Í“D,30d_,_—ÈÉ,
áiØù_Ð+é-70℃|à&Äd。
1.3 \,Ûµ
iíÐ=ŽTÊK A4F6 A4Rè234(
)Y+«èöYáM
。3'?N©ÊKT7dT18n»
^CËÐ+
。5'?#BÊKNUP6UPMnBÜß
¶ÜÊK

‘ýÊKÏdDNAMANÕÖ6Prim
erPremier5.0ÕÖ\‘²²(g1)。
`1 ÝÞߌ犕狑犃犆犜2Nø2ÌÍ\,
Table1 Primersusedforcloning犕狑犃犆犜2cDNAfrom
犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng
ÊKïS
Primer
name
ÊKlj
Primersequence(5'----3')
ÊKŸA
Sequence
length/bp
A4F CCGTTCTGTCCTTGTATGCCA 21
A5R CCACATCTGCTGGAAAGTGC 20
T7dT18 ACGACTCACTATAGGGC(dT)23 40
oligodT12
ACGACTCACTATAGGGC
TTTTTTTTTTTT
29
GSP3' CGGCCCTTGCTCCTAGCAGTATGAA 25
GSP5' CCACGTAAGCAAGCTTCTCC 20
UPM
5'CTAATACGACTCACTATAGGGCAAG
CAGTGGTATCAACGCAGAGT3'
45
NUP AAGCAGTGGTATCAACGCAGAGT 23
1.5 Nøâã\2äå
j34|-cDNA !ßànÂP,A4FnS
ÝÊK
,A4RnœÝÊK\‘RTPCR,ií®5
TÐ=Ž

uŠN]
:cDNA1μL,A4F6 A4R
Ü1μL(10mol·μL
-1),Mix12.5μL,îddH2O
Í25μL。uŠÍÖ:94℃ S<4min,94℃<
30s,57℃ïn45s,72℃Áo1min,30MGH_,
72℃Áo10min。RTPCRií¤Kd2%aô¿
ij`b×Ã

ddPEASYT1/N,\‘Ãl。
1.6 3'犚犃犆犈
}Ïþ—kTÐ=lj²²déií®53'?
TÊK GSP3'(g1),·_Ïd A4F/T7dT186
GSP3'/oligodT12\‘OâPCR,ií®5T3'?
lj

!ß;PCRiíuŠ:j34|-cDNA
!ßànÂP
,A4FnSÝÊK,3'?#BÊK
T7dT18nœÝ\‘PCRií,PCRuŠN]n:
cDNA1.5μL,A4F 6 T7dT18 Ü 0.5μL(10
mol·μL
-1),Mix12.5μL,îddH2O Í25μL,
PCRuŠÄln:94℃ S<4min,94℃<30
s,57℃ïn45s,72℃Áo1min,30MGH_,
72℃Áo10min。!å;PCRiíuŠ:j!ß;
PCR¤KnÂP,GSP3'nSÝÊK,3'?#BÊK
oligodT12nœÝ,\‘ií®53'?TPCRuŠ,
25μLPCRiíuŠN]:!ß;PCR¤K1μL,
GSP3'6oligodT12Ü0.5μL(10mol·μL
-1),Mix
12.5μL,îddH2OÍ25μL。uŠÄln:94℃ S
031
!1# À8}¦§?:34|-9!ó¦®5(MwACT2)dgW=•
<4min,94℃<30s,63℃ïn45s,72℃Áo
1min,8MGH;94℃<30s,58℃ïn45s,72℃
Áo1min,25MGH_72℃Áo10min。PCR¤
KÀ2%aô¿ij`b×Ã,‡jÊ_dd
PEASYT1/N,\‘Ãl。
1.7 5'犚犃犆犈
}Ï3'RACEkdT3'?ljTÐ=Ž,²²
ÊKGSP5'déií®5T5'?lj,ƁBÜß
›œT5'?#BÊKUPM6NUP\‘OâPCR,
ií®5T5'?lj。!ß; PCRiíuŠN]
(25μL):cDNA1.2μL,A4R(10mol·μL
-1)0.5
μL,UPM(BÜߛœ)2.5μL,Mix12.5μL,î
ddH2OÍ25μL。uŠÄln:94℃ S<5min;
94℃<30s,72℃Áo3min,5MGH;94℃<
30s,70℃ïn45s,72℃Áo3min,5MGH;94℃
<30s,58℃ïn45s,72℃Áo3min,25MG
H
;72℃Áo10min。!å;PCRiíuŠN]
(25μL):!ß; PCR ¤K0.1μL,GSP5'(10
mol·μL
-1)
6NUP(BÜߛœ)Ü0.5μL,Mix
12.5μL,îddH2OÍ25μL。uŠÍÖ:94℃ S<
4min,94℃<30s,58℃ïn45s,72℃Áo
1.5min,30MGH_,72℃Áo10min。PCRií
¤KÀ2%aô¿ij`b×Ã,‡jÊ_dd
PEASYT1/N,\‘Ãl。
1.8 ÑY>rfæçè
lj=•ÏdDNAMANÕ֜’,` mP#
ÏdDNAstarÕÖ\‘。
1.9 ,éê÷>r
ljT@”¸D6 ORF¿Qè NCBIT
BLAST Äl\‘ (http://www.ncbi.nlm.nih.
gov/blast/)。ÏdLf=•ÕÖExpasy=•®5
³)T…®WT=m`6?`­

ÏdInter
proscanLf=•ÕÖD…®WljÐ=ep‘=•

<=Lf=•ÕÖ TMHMM \‘¸e
pSÃ

…®Wl j T < Ï Q  è expasy
protscaleSÃ,å9epÏdLf9KÔ:+ÕÖ
GOR4\‘SÃ。Ïd DNAMAN ÕÖD…®W
lj\‘•¸DƐ­]^\ð{

1.10 г1犚犜犘犆犚>r
Ð34|-Gm“D30dkdÈÉ(Òâ@1.
2)。²Âs),€Û,š‰?3M6@ÓùÍÖ。€
ÛÓù2

ÐÈÉ>L4℃“Dà&&Óù6h,àr
f:

)ú5

š‰ú5Óù2

ÈÉu!ÍÙ&f
O

Ð}]=¾ºL200mmol·L-1NaCl,20%PEG
6000Óù6h,àrf:,þ.}‡ýáiØù_Ð
+é-70℃|à&Äd。À_=¾›fcRNA(Ò
â@1.4),Æ\‘`b(RNA T×Aß#),j?
`TRNA tÚNncDNA(Òâ@1.4),Í&f?
`

ÏdA4F/A4RÊK\‘RTPCR(Òâ@1.5)
uŠe5_

f?`TPCR¤KÀ2%aô¿ij`
b×Ã

jÍÜT3AD©®5gWT–o

2 pqW>r
2.1 犕狑犃犆犜2âã\…ëZ[pq
j34|-!ßàcDNA nÂP,ÏdA4F/
A4RÊKií,kd1͟An656bpT®5´
,

ë1),ÀDNAMANÕÖ\‘lj¸D"4,
.Mljþ°gT34|-9!ó¦®5
[18]
Ð
=´,œà&š

Ú34|-9!ó¦®5ß,Ð
=Žlj

=1 犚犜犘犆犚Z[犕狑犃犆犜2Nøâã\æì=
Fig.1 ElectrophoresisofRTPCRamplification
productof犕狑犃犆犜2conservedregion
ü
:M:DNA=m`FÏDL2000;
1:犕狑犃犆犜2®5Ð=Žií¤K
Note:M:DNAmarkerDL2000;1:Amplificationproduct
of犕狑犃犆犜2conservedregion
2.2 犕狑犃犆犜223'犚犃犆犈pq
}ÏÐ=Žlj²²8ºÊK GSP3'ií
犕狑犃犆犜2T3'?lj,d A4F/T7dT186 GSP
3′/oligodT12\‘OâPCR,!ß;PCRïÍÜ,
!å;PCRkd1͟An422bpT´,(ë2),
ÏdBLAST\‘¸D°P,Y´,GJK9
!ó¦®53'?lj@”ûs(>90%),UhÚ
犕狑犃犆犜2®5T3'?lj。
131
! " # $ !24"
2.3 犕狑犃犆犜225'犚犃犆犈pq
}ÏÐ=Žlj²²8ºÊK GSP5'ií
犕狑犃犆犜2T5'?lj,jSMARTRACEcDNA
Amplificationkitš’T!ßàcDNA nÂP,Š
dA4R/UPM,GSP5'/NUPÊK\‘OâPCR,
!ß;PCRïÍÜ,!å;PCRkd1͟An
790bpT´,(ë3),ÏdBLAST\‘¸D°P,
Y´,GJK9!ó¦®55'?lj@”û
s
(>90%),UhÚ犕狑犃犆犜2®5T5'?lj。
=2  犕狑犃犆犜23'犚犃犆犈Z[4,
Fig.2 3'RACEamplificationproductof犕狑犃犆犜2
ü
:M:DNA=m`FÏDL2000;1:3'RACEií¤K
Note:M:DNAmarkerDL2000;
1:Amplificationproductof3'RACEof犕狑犃犆犜2
2.4 ÑY>rfæçè
   Ï d DNAstar Õ Ö D Ð = Ž、3'RACE、
5'RACEkdTef\‘P#,kd1͟n1506
bpTlj,³»œWTPolyAep,ýïn34|
-9!ó¦®5犕狑犃犆犜2。
=3 犕狑犃犆犜25'犚犃犆犈Z[4,
Fig.3 5'RACEamplificationproductof犕狑犃犆犜2
ü
:M:DNA=m`FÏDL2000;
1:mb½;2:5'RACEií¤K
Note:M:DNAmarkerDL2000;1:Blanklane;
2:Amplificationproductof5'RACEof犕狑犃犆犜2
2.5 ,éê÷>r
ÏdNCBIORFFINDERÊQ‘¼>â(É
ù

~ë41˜,Ylj»J1131bpT ORF³)
376M…®W,Æò»J5'UTR129bp,3'UTR
246bp。
=4 ÝÞߌ犕狑犃犆犜2Nø2犗犚犉>r
Fig.4ORFanalysisof犕狑犃犆犜2from犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng
  ~ë51˜,BlastXefgh,犕狑犃犆犜2Ý
a¢ ë u ^
(犜狉犻狋犻犮狌犿 狌狉犪狉狋狌)、å v C R -
(犅狉犪犮犺狔狆狅犱犻狌犿犱犻狊狋犪犮犺狔狅狀)、‡m(犛犲狋犪狉犻犪犻狋犪犾犻
犮犪)?ÞJtsT@”(>98%)。
ÏdLf9KÔ:+=•ÕÖ Expasy(ht
tp://web.expasy.org/compute_pi/)=•gh,Y
231
!1# À8}¦§?:34|-9!ó¦®5(MwACT2)dgW=•
®5³)ó¦T=m`n41.606KDa,?`­n
5.29。ÏdInterproscan(http://www.ebi.ac.uk/
Tools/pfa/iprscan/)Lf=•Õ Ö D 3 4 | -
犕狑犃犆犜2®5T…®Wlj\‘Ð=epS
Ã

efghY…®Wlj¬éActinˆ‰ó¦,Æ
ò»JActinÐ=Ÿ­(ë6)。
=5 ÝÞߌ犕狑犃犆犜2Nø犗犚犉¯NqÑY犖犆犅犐<2‚$pq
Fig.5 Blastresultsoftheputativeaminoacidsequenceof犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿
Keng犕狑犃犆犜2openreadingframe(ORF)inNCBI
=6 ÝÞߌ犕狑犃犆犜2Nø2âãpÝ]>r
Fig.6 Conserveddomainanalysisofthe犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng犕狑犃犆犜2
  ÏdDNAMANÕÖD34|-、»z、œá
âºu
(犛犪犮犮犺犪狉狌犿狅犳犳犻犮犻狀犪狉狌犿)、6¾(犜狉犻狋犻犮狌犿_
狌狉犪狉狋狌)、Ý a ¢ ë u ^ (犅狉犪犮犺狔狆狅犱犻狌犿 _犱犻狊
狋犪犮犺狔狅狀)、åvCR-(犛犲狋犪狉犻犪犻狋犪犾犻犮犪)、‡m(犎犲狏
犲犪犫狉犪狊犻犾犻犲狀狊犻狊)、Sj{(犞犲狉狀犻犮犻犪犳狅狉犱犻犻)、çT
(犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪)、&ZA(犃狀犪狀犪狊犮狅犿狅狊狌狊)、
UC
(犕犲犱犻犮犪犵狅狋狉狌狀犮犪狋狌犾犪)6ìí12MJKT…
®Wlj\‘•¸D

°P Actin…®WljÞ
JtsTÐ=

ë7)。…®Wlj]^\ð°P
34|-Actin…®WljÝa¢ëu^(犜狉犻狋犻
犮狌犿_狌狉犪狉狋狌)T Actin…®Wlj<<‰]Ýä

ë8)。
D󦸝TSÃJÅé‡"ù”ó¦Te
p6¦§

ÏdLfÕÖTMHMM(http://www.
cbs.dtu.dk/services/TMHMM/)DY®5³)T
ó¦lj\‘SÃ

efghY®51³)Tó¦
F6+L¸ep

¬éȸó¦

ë9)。
ÏdLf9KÔ:+ÕÖExPaSyTProScale
(http://web.expasy.org/protscale/)D34|-
犕狑犃犆犜2®5³)Tó¦\‘</ÏQ=•,k
d~ë101˜T<ÏQ=>éf。‘&0jS
ëogÏQ
,0jœëog331
! " # $ !24"
=7 ¯NqaÑY‚$pq
Fig.7 Multiplealignmentresultsaminoacidsequences
ü
(Note):犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng:34|-;犣犲犪犿犪狔狊:»z;犛狅犾犪狀狌犿犾狔犮狅狆犲狉狊犻犮狌犿:œáâºu;犛犪犮犮犺犪狉狌犿狅犳犳犻犮犻狀犪狉狌犿:6¾;
犜狉犻狋犻犮狌犿_狌狉犪狉狋狌:Ýa¢ëu^;犅狉犪犮犺狔狆狅犱犻狌犿_犱犻狊狋犪犮犺狔狅狀:åvCR-;犛犲狋犪狉犻犪犻狋犪犾犻犮犪:‡m;犎犲狏犲犪犫狉犪狊犻犾犻犲狀狊犻狊:Sj{;
犞犲狉狀犻犮犻犪犳狅狉犱犻犻:çT;犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪:&ZA;犃狀犪狀犪狊犮狅犿狅狊狌狊:UC;犕犲犱犻犮犪犵狅狋狉狌狀犮犪狋狌犾犪:ìí
=8 ÝÞߌ犃犮狋犻狀Wgí+,
犃犮狋犻狀st2)*ÒQÓ
Fig.8 Phylogenetictreeof犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿
KengActinwithotherplantActins
=9 犜犕犎犕犕îO$ÝÞߌ 犕狑犃犆犜2
GE¯Nq2ï²]ð²
Fig.9 Predictionoftransmemberanceregion
ofthe犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿
Keng犕狑犃犆犜2aminoacid
sequencewithTMHMMsoftware
  ÏdLf9KÔ:+ÕÖGOR4(http://npsa
pbil.ibcp.fr/cgibin/npsa_automat.pl?page=
431
!1# À8}¦§?:34|-9!ó¦®5(MwACT2)dgW=•
npsa_gor4.html)D34|- 犕狑犃犆犜2®5³)
T…®Wlj\‘å9epSÃ

~ë111˜,α
;Ø25.53%,β´’Ø25.80%,ï£("éØ
48.67%。
=10 ÝÞߌ 犕狑犃犆犜2NøGE
st2ñòóÏ>±=
Fig.10 Thehydropathydistributingof
犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿Keng犕狑犃犆犜2aminoacids
2.6 c~z{$犕狑犃犆犜2Nø`Ôµ³Ï>r
“D30dT34|-ÈÉj6@ú5(s)、
€Û

š‰

Óù6h_›f RNA,\‘ˆ©`RT
PCR=•gh 犕狑犃犆犜26ce÷tlú5TU
V

ÜGÓùÍÖ%gW`غ

ë12)。g
h犕狑犃犆犜2gWû¨©,6ce÷Hlú5。
3 ˆ‰Wp‰
  9!ó¦Ú—¨9KH.Dú&T®»epä
=

‘§ñ’ó8T?VDúep

L=mQRS
žû•«T9ù9ð=Ä

³´Ô{+"

.E
.Fcdj|TGHÏd

®5ÚNb
、mR
NAîÃ6ûn?,5.,‘ÞJ•«T/0M

[2021]。
jJK9!ó¦nÌñ’T!\?VDú
]^þ’nJKH.9K+/0þTp­ã
ß
[22]。
»/0Œ“—k¡34|-9!ó¦®5
àŸcDNA,ëì¡?-9!ó¦/0T“ý˜,n
=•?-=m9K+/0›œ¡®¯‡ý

=11 ÝÞߌ 犕狑犃犆犜2NøGE¯NqÑY2 ºpÝð²
Fig.11 Thesecondarystructurepredictionof犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿 Keng犕狑犃犆犜2aminoacidssequence
=12 c~z{NOPÝÞߌàÇ
stNø
(犕狑犃犆犜2)2`Ô
Fig.12 Theexpressionlevelof犃犵狉狅狆狔狉狅狀犿狅狀犵狅犾犻犮狌犿
Keng犕狑犃犆犜2underdifferentstressconditions
  2/0wx4n,34|-ÚßGúŽ‰6
ø¥Tp9q-

µvYKGþ/jn¬å9?
^W°JK6ž3ÐÑT(cK¾9ä[19]。
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,
[1819],
»BCLjS“ýT®¯S

Í34|-&
d—k¡ 犕狑犃犆犜2®5,Y®5cDNA àŸ
1506bp,³»1M1131bpT¼>(Éù,»J
ActinÐ=Ÿ­,ÚActin¬T’¯。Y®5§
³)376M…®Wä’T9!ó¦,³)ó¦T=
m`n41.606KD,ùy?`­n5.29。Yó¦¬
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,α; Ø
25.53%,β´’Ø25.80%,ï£("éØ48.67%,
.}ëÏٚJK9!ó¦®5ljÜG8
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Y®5Ýa¢ëu^
(犜狉犻狋犻犮狌犿狌狉犪狉
狋狌)、åvCR-(犅狉犪犮犺狔狆狅犱犻狌犿犱犻狊狋犪犮犺狔狅狀)、‡m
(犛犲狋犪狉犻犪犻狋犪犾犻犮犪)?ÞJtsT@”(>98%),g
hY®5L\ð=Ä&gPÈPÐ=

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”•AB
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