免费文献传递   相关文献

Construction of cDNA Library and Analysis of the Expressed Sequenced Tags (ESTs) Characteristics of Dormant Seeds of Davidia involucrata

休眠期珙桐种子cDNA文库构建及EST分析*


以休眠期的珙桐种子为材料,利用SMARTTM cDNA Library Construction Kit构建珙桐种子的cDNA文库。结果表明: 该文库的滴度为1.5×106 pfu·mL-1,重组率为98%, cDNA文库插入片段大小主要集中在0.5~1.0 kb。随机测序获得148个EST序列,拼接成127个uniEST,经网上BlastN及BlastX分析,其中75个序列是已知功能的基因标签,相似性较高,而32个序列在GenBank中相似性较低,13个序列在GenBank中没有匹配。这些数据有助于对珙桐种子的相关功能蛋白及分子机制的研究。

A cDNA library was constructed with dormant Davidia involucrata seeds by using SMART cDNA Library Construction Kit. The library had a titer of 1.5×106 pfu·mL-1, with recombinant rate of 98%. The inserted cDNA fragments ranged mainly from 0.5 to 1.0 kb. Total 148 ESTs were obtained from randomly sequencing, and 127 uniESTs were obtained by contiganalsis. By searching on line with BlastN and BlastX, 75 uniESTs were determined to be the tags of genes with putative function and high similarity, 32 uniESTs had low similarity and 13 uniEST had no significant similarity to any protein or DNA sequence in the databases. The present ESTs provide a valuable resource for further research on similarity protein function and molecule mechanism of Davidia involucrata seeds.


全 文 :! !"" ! #$#
% $ $ &$ #$ %
& ’ ( )
’()*+,)- ’)./-* ’)+)(-*
/012!"*+02#$
3456*
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
% $ $ &
56#qr1{ 47+-s7M{A *’,@B"
’ $#$% ()*% ,+S8 ,-G8
F#6x>z9*)X=())H ,& 98:$$%T%6T1wXY)aZ[bctu\]^_‘ B‘ 9%#$$$T
8 6B‘z9)H@{X;)aX;LK\]^_‘ B‘ 9%#$$$f
! "# Y56#~qr1{gyX*>$ ’;-<,,; 47+- .=>?@?A (0BC5?D45=0B E=5M{qr1{~ 47+-s7PQ
RSTUos7~h,g #2" F #$9 GHDHI.J #*\DCg &KL*47+-s76u3W*0NO}7D $2" M #2$ N>P4
s8;M* #!K- *’,;c*s/m #%:- DB=*’,*Xs" O1@C5+A O1@C5P@B*B7 :"-;cXBV<=~jz
/t*ZCHc+*G 8%-;cD QRBO@BN7ZCHcf*#8-;cD QRBO@BN7#bujP45|}b-[mq
r1{~Zl<=_}A@{sL~QRP
$%&U qrT1{T56T47+-s7TSW;c/t
’()*+#’:#K2!9 ,-./0#- ,12+##$$# J :!KK3%$$&4#$ J $$9& J $"
|&’#U%$$K J $K J $9P
j()*UGU+4()j(F8$9:$%$&fP
"Kv_g01?=P
!"#$%&’(%)"# "* (+,- .)/&0&1 0#2 -#031$)$ "* %45 678&5$$52 95:’5#(52
;0<$F69;$f!40&0(%5&)$%)($ "* +"&=0#% 9552$ "* !"#$%$" $&#’()*+","
S= Q@BT#*% ’D UV=W=@B% .= X=BT5@08 F# 6 !"#$$% $& ’(&) !"()*")+*,#(*- .)+/ 0$12-% 3*(4)1+(/5 0-*"#$*6 98:$$%T
% 6 7)5 ’-8$1-/$15 $& 9"$%$65 -*: 9*4(1$*2)*/-% ;1$/)"/($* <)"#*$%$65 $& !("#=-* ;1$4(*") >(-*5-*6 9%#$$$T
86 7)5 ’-8$1-/$15 &$1 >$%)"=%-1 ?($%$65 -*: ?($@#-12-")=/("-%+ $& >(-*5-*6 0$12-% 3*(4)1+(/5 >(-*5-*6 9%#$$$f
-/$%&0(%U - 47+- 1=>?@?A Y@C 40BC5?D45RZ Y=5V Z0?I@B5 A-4(:(- (*4$%="1-/- CRRZC >A DC=BT ’;-<, 47+- .=>?@?A
(0BC5?D45=0B E=5 6 ,VR 1=>?@?A V@Z @ 5=5R? 0H #2" F #$9 GHDHI.J #*Y=5V ?R40I>=B@B5 ?@5R 0H &KL 6 ,VR =BCR?5RZ 47+- H?@TIRB5C
?@BTRZ I@=B1A H?0I $2" 50 #2$ N>6 ,05@1 #!K *’,C YR?R 0>5@=BRZ H?0I ?@BZ0I1A CR[DRB4=BT*@BZ #%: DB=*’,C YR?R 0>5@=BRZ >A
40B5=T@B@1C=C6 OA CR@?4V=BT 0B 1=BR Y=5V O1@C5+ @BZ O1@C5P*:" DB=*’,C YR?R ZR5R?I=BRZ 50 >R 5VR 5@TC 0H TRBRC Y=5V GD5@5=\R
HDB45=0B @BZ V=TV C=I=1@?=5A*8% DB=*’,C V@Z 10Y C=I=1@?=5A @BZ #8 DB=*’, V@Z B0 C=TB=H=4@B5 C=I=1@?=5A 50 @BA G?05R=B 0? 7+-
CR[DRB4R =B 5VR Z@5@>@CRC 6 ,VR G?RCRB5 *’,C G?0\=ZR @ \@1D@>1R ?RC0D?4R H0? HD?5VR? ?RCR@?4V 0B C=I=1@?=5A G?05R=B HDB45=0B @BZ
I01R4D1R IR4V@B=CI 0H A-4(:(- (*4$%="1-/- CRRZC6
>51 ?"&2$U A-4(:(- (*4$%="1-/-TCRRZTZ0?I@B4AT47+- 1=>?@?AT*’,C
qrFA-4(:(- (*4$%="1-/-fJ2|8*I=w\*
K!Ufw12:;nK~xp1*b,.;F-W
8*g_G6Z1~@b:;*>cgGU2}\]
bc:;*e@$~~+2,b6r:;*>yg:
;@~,*z{-FKv_(*#&&&T.= )/ -% 6*%$$8fP
qr34@ED2Tx$,x$dxbxxT1x2,
AY| :w*@E9:c}*,=}cI*JR[2
5Yq(nqr1{~56#c0*2? % M 8 $,=O*
qr1{~,+56H-X0#g:&’Z[G%m
~X6XY@HPghQRB56~sL*"Jg
)*B;R^%YQMF"#$*#&KKTZ(1(*
#&&8T[eU*%$$%fxRf;)DmF%*D(*
#&&"fxfXSR;SFR&.(*#&K&TY’)(*
#&&KTd()(*%$$8T)QH(*%$$!T"*(*
%$$"fxD2NU(*%$$8Tr+(*%$$8f(E1Q,mqr1{~
56A_AGHoph*}~<_QRT4GD@
{bd"q_}Syjzvd~QRTXQ}P9
QRY56#~qr1{gyX*M{GR?! #
-qr1{ 47+-s7*opSW;c/tF*’,f@
B*Y#QRBg:Z[~Zl~<=_}Ajz*
9g;@{|b"QRBsL]OQ}P
# yXavv
@A@ @A
qr1{[ %$$:$ #%%#+T1,t+4b
cnU{-.&/‘~}+T*BR0[‘wb‘P
!"# }c
t$ !"#$%%" &’(# )*+,-,. /0123,4&3*01 5*3F1
+ /60137&8UcIg !"#$%&’#$’( #)*’ =)>?@647F1+ /60137&8 U
!.237DF1+ A,0D7C- UoM@BaP
!"$ <=
>FGF> JK"LM $(#N’ #$Nn~ )*/6v
]‘! $(#F)* %+ (* H*9b^*Qp7?D>I D*1[*@u KLAMA*Q\T
X29|iT29b NII")78~3]$!^
FO D06H)P > )*/6*Dt$3@uB[ P ~ KL#P4j!"*>I DD06H)
P >’5L-y +H AMA*FR D);tO7*&@[
F[ K QKT6P‘lT6[~;tO7i;t
F#R 2fP0":^|iT\2+v~>FR D);t
O7*;t GI D*1*O":^P$ SIL !"F
P I D*1*n89+[JRI")
+ i ^ 7F >FKL !’!*>RI DD06H)P > (-/6*
RI DD06H)P > :’%#*>I DD06H)P > %,*2?T/6*9TSFR*
F9TSFOf*G) U:; Us<"Ff*:; Us<
"Ffy:;3]Prf3]3*0bsZy
$(#+^&@[F[;t>R D*1*‘":^P")
[ >f3]~":^7@u >V>I qe~K D06H)P >
=->y <,qe~8!"FK Qf*P GI D*1P[E;;tGI D*1*B":^*"89@u
P >=->89 $(#P[E;;t
>I D*1*$ SRLy OIL8!"u89**+89[+
[ >G III ,HD*1P >P‘ >")uU$[@33,qa,
yc,i8*‘ G")$[5I%OM\z@BP
>FGF< &’(# OPQ3 l} !"#$%%" &’(#
)*+,-,. /0123,4&3*01 5*3 ^_.C5TM{ &’(# s
7P‘G") $(#g22*$ /’!$VGW A/$ A,*D7, y
!"#$% XM%" Y6*C014&6703*E7*9|zEm6? &’(#P
$$ RW A/$ A,*D7,y /’!$VGW A/$ A,*D7,0TH;
VWEmg? &’(#P4[XT_}" 5 K;*
,-’%"B*$X /8,0D- !9*1?KII @ui*@;9|
}*[ RII +9 &’(#3WPo3WaEq!%,*96:;<
~OPA%/[*t$ A-&B-C717 )-D+E- ’(#
A-&B-C*1C !.237D opq?8&*z2[S*Bcq
^i^@&[>FR D);tO*[ P OI Qb‘*qg
M{~qr1{~ &’(#s7P
>FGFG OP%HI;R0ST ; &’(# s77
‘lR")Bcq^i^*jo > U >I ,B*4[‘
>")aI DD06H)
P > "C!YK \t~T6N<
~ ! H #)*’ =)>?@647 [E*GS Qij>R D*1*a< D)
’;~"b5I[F*moD GS Q71~F)@V
"C!YKfb2"*GS QN< J Z >I 8*l}BcC-
|*qrs7h,Pl} &’(#s7h,*‘g}B
cq^i^@Xjb XA%[FRI"CHD)
P >fy =?C-6
FRI"CHD)
P >f~"b5I7*GS QN< O Z >< 8*q
|b2"%m~%Cy}C|**qrBcq~DCP
>FGFK &’(# OPUVjWST 4s‘‘Bc
C*+[>II") > \!,B F^"C/6< IFI> D06H)
P >*
%,*2?T/6 9T SFO*IFIGR D06H)P >*TM IFI>L*B;{
|IFI> D06H)P >fP‘R")?g22*t$!%,*96:;<
5*3]O~ RW P y GW P v]tH;op A/$z2*
$ >L5I%OM\zi8 A/$3W~*0*qL
i8l &’(#s776u ’(#3W~0,*0P
>FGFR OPXv $X A/$i8jb‘HgD~
Bcq|; ! H #)*’ @"K*\DBcq ’(# Z;mS)*]‘S)*|;
! H #)*’ ’TR&*b‘|;{P
>FGFJ ’YT.2;< Y"{H;FRW P
/%//[#[#%/%[[#/[#[/ P GWf*K{H;FRW P
%##%#/[#/%/#/%#%#[[[ P GWf;T"+vEX;
bRU<8;P:-X;@A ’(#!%#$ y @*0
:E*3*08;S*X~;cops/*BkEqxH
;;c[*@/t$ @6-23( y @6-23= iv (-3*01-6
/7137, ]0, @*037&81060C. X1]0,D-3*01F(/@Xf~XF-y
_}S|}7*op;c~h=H@BP
< QRa@B
#"! ABVVC %&’P8;H
+S}~ $(#XM{ &’(#s7~l^*‘ G
")! $(# op >F>L~5I%OM\zi8Fm
>f*ST O! ,$(#\z52Zu:G*2s,#g >O!~ <,P‘ >")uU$[@33,
qi8*8* Y’*5TS]‘~! $(# S}c+*Lg$[
&’(#s7~M{P
#"# ()&’,DEy9P8;H
0abHH2v~ &’(# aEq!%,*96:;< %
/*_}8&[|;*I{c ! F #)*’ =)>?@647PD
jb XA%[y =?[-6 ~b2"LqBcC|Pqr
*l*S{ &’(#s7h,g >FR \ >IJ 9]4HD)P >*X
IS & ’ ( ) KR"
%Cy}Cq|*!"#$s7~\DCWX %&’P
4s‘‘}+BcC () -*>$ *+, tui
86u "#$3W~*0PQRST*6u "#$ 3
WNO}7D )-. / (-) 01WRFm 2fP
m ( qr1{! ,#$
3456( 789:; ,#$ 8< !"#$%$" $&#’()*+"," =>>?=
m 2 \zi8X *+,z2l~gD{6u "#$3W*0
34562 @>;>;>!9A8BC8A>=4= 8< *+, :DB;4<4!:948E 8<
9C> "#$ E9= 4E=>A9>? 4E B;:=D4?= $&#’()*+"," =>>?= !"#$ ;41A:AF
GU"H2))) D:A0>AT( I ()UX *+,z2lS)7
"#$6u3W *+, :DB;4<4!:948E 8< 9C> "#$
E9= 4E=>A9>? 4E B;:=D4?=6
!"# $%&H4945)T
4s‘‘ (.)-BcC*8;M* (J&- KL7;
c*XT KL7;cbLs/[vM* (2M- KL7;
c*K! KL7~ &.-&’*;c0,g %N2 / ( &%( 1B*b
y0,g( JO2 1BP>$ #+PQ~ R>! L!A>>EU;~/
9BkEq>?~;c*Bk0[ ()) 1B ~;c M
5*K6y8;}~ J-M’PXTs/yBkEq;
c[*M*bL;c (2)5*K! KL7~ &(-(’*;c
0,g ()N / ( M(& 1B*by0,g .2N 1BP
0T P;:=9Sy P;:=9#@A (2)5bL KL7;c
@/D #+PQ"~kJK_}S|}7YAX-;
c|}77opKv@B*QRST*vb M. 5
KL7;cD_}SW=XF-|b"abh=;
c*K!|~ O2-.’*B7 225g<=HV_}*K
(&-N’*b N2 5 KL7;c0TVmZCHcf*K
(%-2’*(N5 KL7;c!uj*K (&-N’FS (fP
m4 .N5 KL7;cjoh=VmQRopX
;)<=@^L*D@g & F^+L(*2)).f*89
$}SRx_}SEmx|zx=}SRx*hQMx
>IsCxAM[[yBC^P9s<_QRHm
NPB7_}SEmZl~;cSKVL)E
F2%-N’f*BfX>I|CF(J-O’fy*hQM
F(J-O’fP*E| KL7 LD2!:;(,a
F-+".$%’/0$0 ,1"($"&"f7_Xh=;c*aWuj~
<=_}7bqr1{7Nj~E%_}xm!_
}*YA1{7aZ[g:HZl~<=_}Pq
r1{y@<=_}mVQRHS (P
m N .N-qr1{ KL7;c~X;)<=@^
3456N 3TE!948E:; !;:==4<4!:948E 8< .N KL7= 8<
!"#$%$" $&#’()*+"," =>>?
N Qca]c
]‘+S}~ ,#$*XM{ !"#$s7~j!
yl^P)[qr1{7j*}~E%yE)*Y
pM*+a,+=}~ ,#$*9QRm H4+;v]‘
! ,#$opNoM*h+S}~! ,#$*Z6g$
[ !"#$s7M{~O1Ps7~h,x\DCA
6u3W~*0X%t !"#$ s7S}~\O.
/P2?g5*s7~h,=WX(). BqgbL~s7FH4 2, "( 6*(%%&fP9QRM{~q
r1{ !"#$s7h,g (-. U ()O BCg %&’*5Tos7S}c+P
KL7tu>"gX21;jzs77>?M*
jzD7~HV@AjzSWy0w;c~vv
F$?:D= 2, "( 6*(%%(TUZT(*2))OfP9<_SM
*~ KL7;c=a$[M‘B60 !"#$*oGM*
B60jz;cY$[<=QRP0Tmqr1{
!"#$s7~ KL7 8;A;cVm@B*b NM-.’
KL7;cD|}77~ZCHcfW!uj*L=
Xwjz*JJ-2’KL7;ch=Hc+**Xh25
<=jzP
(M! ()# = H(U56#qr1{ !"#$s7M{A KL7@B
C ! "#$%&$’’U5A>?SFPNO
(&)* ! +,-.&/01,$ ,2 3,-,4,56 7083 29$:80,$ 5#$#1 0;#$8020#; 0$ "#$%&$’
gDI
!"#$% &#’
0,
(%$)*+,-.
88_}
/0*1*23% 24%$*25261*2#$
h=;1
!#7.18%4 9.%62%9
@7
:6#8%
ZCH
;4%$*2*<,=
>?@AAB CDE
N;{Qr8._}
!1F G H4%.%$4%$* 7%7-81$%H-2$42$) .8#*%2$ 1$$%I2$
(,a
!"#$%&’()%) *+#,%#-# JKC EC
>?@ADC CEJ
lX|eIWz{ FL^_}
M0N18<#*26 *81$9"1*2#$ 2$2*21*2#$ 516*#8 FL 5172"< .8#*%2$
(,a
!"#$%&’()%) *+#,%#-# OAP EE
>?@AFD D AKA
G)q%MiO"&E /_}
Q2*#6+#$4821" )"<62$% 4%618-#I<"19% 6#7."%I /H.8#*%2$
^s(‘
.’(/,/) *"01/,’%&0) KCP BC
>?@AFO EP 1{m!_} /QJD :%%4 71*081*2#$ .8#*%2$ /QJD *j 2,34%-0 1#5 DKA EC
>?@AFP OPO E:7_} E: )"#-0"2$

>?@ACP DOB 3hL-_}" R9.18*26 .8#*%19% 8Q 7#-%+’* 0)4/,0-*# DAC BE
>?@AJA DAA
4AZl]RqHD
S-2T02*2$H8%"1*%4 7#4252%8HD
|r
8"39# )#*%:# ;$4261 >8#0. DJE EC
>?@AJC FPC
STjzZC_} J*G)q$#%!
&25SH"2N% .8#*%2$ J*72*#6+#$4821" .8%6089#8
(,a
!"#$%&’()%) *+#,%#-# CKK EB
>?@AOC DOE i|A U+$D >Z 6’,#-/1 4’110")’-%% PCVK KP
>?@AOJ FKO
G)qRyi("
Q2*#6+#$4821" 2$$%8 7%7-81$% *81$9"#619%
UVWX
;+,#13&’1’-#) "0%-+#"&*%% FEB PA
>?@AOB DOP
+SetUt DK2FO-etmx P_}
:#"0*% 61882%8 5172"< DKF7#$#618-#I<"26 1624 *81$9.#8*%89f*7%7-%8 P
CfY
<#-%’ "0"%’ DAK JP
>?@AKC EF
*h+A -H6D&Eq BF4A*h+A;V"&Eq PVA N0_}f
!<*#6+8#7% -H6D 6#7."%I 90-0$2* BF S-2T02$#"H6<*#6+8#7% 68%406*19%
6#7."%I PVA N0 .8#*%2$f
fZQ
6’,#-/1 */$0"’)/1 BAVB EK
>?@AKJ DOO
15g_} DP
W%1* 9+#6N .8#*%2$ DP
.[E
;’&’-’()%) ,#-40’,#*# DPE KO
>?@AEJ DAF
\)%]t (<9QQMz%L_}
/%.*24#)"<61$H-2$42$) (<9Q 4#712$H6#$*12$2$) .8#*%2$
(,a
!"#$%&’()%) *+#,%#-# DFD KC
>?@AEO D BDB (Z*H_} X65D .8#*%2$ >?@AEP CDF $%n:m@]t_} :068#9% 8%9.#$923% %"%7%$* -2$42$) .8#*%2$ 89 >%*%) :%-%?0"# DCB JJ
>?@APD FKF
|({_}
?81$9.#9#$ .8#*%2$
|rF=N:1f
8"39# )#*%:#FY1.#$261
60"*2318H)8#0.f
DJB OD
>?@APE CDK ^D?/9^_} Z!1LJF Z160#"%H199#621*%4 1$$%I2$ Z!1LJF I7 @%4’*%#-# *#$#4/1 JEJ EE
>?@ABJ JJP JP N0%_}$# JP N0 )"<6#.8#*%2$ .8%6089#8 rsJ ;’"3,/) #:0,,#-# FOE OE
>?@ABK DCJ R?/
Em" O!j
R?/ 9<$*+19% %.92"#$ 90-0$2*
2o Z
.+3,,’-’1# "/)4%?’,%# PBVJ PP
>?@ABB BB
0XX_}
:71"" $06"%18 82-#$06"%#.8#*%2$
(,a
!"#$%&’()%) *+#,%#-# DEB BO
>?@DAA CFA KA:-HX%q_} /A KA: 162426 82-#9#71" .8#*%2$ /A *j 2,34%-0 1#5 JEK PK
>?@DAO KFK
IQR_‘a1XF-QroREm" KF N0 QM*G)q$#
&RUH4%.%$4%$* 71"26 %$[<7% KF N0 29#5#87*72*#6+#$4821" .8%6089#8F&RUH
QMf
fZQ
6’,#-/1 */$0"’)/1 DPE PC
>?@DAK FCA P5Y\ :|i" >"0*1*+2#$% :H*81$95%819% +b{ 6#,%4’"-%# $"#4+%#*# CDP KK
>?@DDD OPP SQS!
ZC U&R]&Ut_}
SQS!H"2N% U&R 8%.128 5172"< .8#*%2$
(,a
!"#$%&’()%) *+#,%#-# PCVF JA
>?@DDC KC (ZU_}^3 C/\;];OAC Q%*1""#*+2#$%2$H"2N% .8#*%2$ *<.% C /\;];OAC 89 >%*%) :%-%?0"# DAF EF
>?@DDK CPB X%q_} (CR 2^-#9#71" .8#*%2$ (CR I7 @%4’*%#-# *#$#4/1 EAJ BJ
>?@DCC OE fwqC(;{8_} !#"4H2$406%4 ."1971 7%7-81$% .8#*%2$ ciZ 7/)# RLL >8#0. BAVB PB
>?@DJO PB
|zVLz{ Dhtq
?81$9682.*2#$ %"#$)1*2#$ 516*#8 D +#7#"#)
|r’91
8"39# )#*%:# Y1.#$261 >8#0. DJC BD
>?@DJP FOO
*c5g_} R*d.7cSd_} R
/+1)% 9+#6N .8#*%2$ R*/9.R
%{X
;3#-’*+040 9.’ /!! PPAD FFJ JE
FE & ’ ( ) JO"
0T@B*Xhqr1{aZ[Zg:<=_
}~ !"#;c*#15g_}F$%&’ ($)*+ ,-)’%./fxi
|AF0$/1fxfwqC(;{q8_}F*)203./04*%0
,2&(5& 5%56-&/% ,-)’%./fx| z V W z { 7 1
F’-&/(*-.,’.)/ %2)/8&’.)/ 9&*’)- 1 $)5)2)8f(P45_
}VDqrX0~++~xfwz/~Z[7I5
\O?$*+Xqr1{g:BZ[~\O_}P
15g_}XX;~uqxD2x*h(hBX.~
w,9:EA’;fwi"+wi*L#$’Oo
Em~2D_}S*D*hX=.:7I\O?$*
y@15g_};a*h[i|_}‘D;c"~
ZCH*4=m<8efm8~p2*mI+Sh
#*bL*hi|*;G2?hD2~*8HP_e
L(F:;;*=>?{7y !"# @B7m15g_}oph@
B*STBm~hA*DH~?$Pi|Aab+
,~G|H*F=aD:;efOV[#YA^[
+-xfwxd&(9[~:I7*}SWPh:;
*h/SR~bcDfw(9[7E^i*FLY
:k.H/+)j~=X*bc*h8KLP,$
gl[i|A~QROPq>*@/X9[Ki|
Am:;~bc?$HIhL(*:;;;Xfwefi*=j/bc*hA;ef2p~
<=_}P|zVWz{ 7 1X21D*hRmn
‘D/*}SW~E)X%q_}*D*fwy*
2?G(vdL=+I5l^~?$F{E~(*
:;;@fP9QR;*Xh1{m!_}F (%%0
5&’4-&’.)/ ,-)’%./fx4 " 7 * h + A ; V "
F*A’)*$-)5% 63*1 *)5,2%B (464/.’ Cf(*45gJ[q
r1{~o2qQR]Oj!P
)[qrX2w\;1*/,=@g@I*1{
Yp=O*Bmg+,;1*mqr~bc[gOP9QR0TM{qr1{~ *=>?s7*{&
IhBjz,=*mJ[QRqr~KL%;x1{
56=OAg:Z[(]Ohm2~,=y@{X
;)Q}P9QR0T !"#(8;yaBV<=j
z_}h=H@B*mqr1{~@HYAag:
Z[Zl~<=_}oph$q~\]*gJ[o
2qQRB<=_}~@Hs6YAjz~gDS
Ww%] ^ , -
*e)*Y&T*Q{g*( D 1CEF DqrD2N< D:;X6)01*
FGfUGF 7 GGD
Y’)*YHh*331*( D 1CCE Dqr1{be#RT/;;"h
>"~:; Dx,&)H)<*1EFFfU1GF 7 1G%*D*s s*_7B D 1CC@ DqrRf7ixNFA~ !="@B D{#!)<*F:fUCC 7 1;FD
)QH*RJj*/k) D :;;G D:;X0ws1mqr1e;qN<
~KL D$d_)H)<*:GFGfU:C1 7 :CFD
+ L*l @*p m*( D :;;@ D’9nopF /’"$0&1’ 2’$#(%.’fM
q_SW;c/tF!"#f@B Dps)<*F:F1;fU1;G@ 7 1;@:D
_eL*xGH*9so*( D:;;< D~hA *=>?s7~M{y !"#@
B D>d_)<*::FGfUF< 7 G1D
!(X*& k*Yvn D 1CC@ D_G+,@b:;qr~kbABb
c DX;EuH*F GfU:1F 7 ::1D
d()*Kv_*Y=P*( D :;;F D+,.r:;qrR^7~=O
bL;S D:$aZ[X;)<*CFHfUH;< 7 H1;D
R&.*Y/$*-UT*( D1CEC Dqr1{56y=O7-H#-
"h>"~QR D x,&)H)<*CF1fUE 7 1FD
[eU D :;;: Dqr1{~iDQR D$djt)H)P*:;FGfU1E 7 1CD
[eU*U3g*/{T D :;;F DqrAqD2NH)Kv_*EA| D1CCC Dqr1>Xi;9ABKLz{QR DT1z
9)H)Z(1*6Dq*set D 1CCF D+,:;-.%Y$qQRFUfD{
x&’(t*F:fU:; 7 :FD
" **% F D:;;@ DOoqr1{OA~tuQR Dx,qs*FF
F:fUCD
"#$ D 1CEE Dqr1{56~iD)QR D7,&)H)<*EF1fU
F@ 7 FCD
r +*E ,*@]u*( D:;;F Dqr$SN)<*:1FGfUF@H 7 F@ED
{E~*&-H*/ ,*( D :;;@ D*)q77G*OIG)qoR
-il"jz~gDy;c@B Dqrv)’X*:@1FD
UZT*EJH*K5>*( D :;;H D^s$‘+H_A *=>?gD8
;ASW;c/tF!"#(f@H@B D&’()*G:F11fUF< 7 G1D
?0&5( I =*J%22%A K I*L)*&A/% K =*0& ’- D 1CC1 D M)5,2%5%/’&-A =>?
(%N4%/*./8U %B,-%((%0 (%N4%/*% ’&8( &/0 $45&/ 8%/)5% ,-)O%*’ D
"*.%/*%*:@:U1H@1 7 1H@HD
P. #&.Q4*R.&/8 K.&/$&.*P.4 S4.A4D 1CCE D M)/(’-4*’.)/ )9 *=>? 2.6-&-A )9
($-.5, 30(’0*+ .)%(0(+%+F M-4(’&*%&U=%*&,)0&fD ?*’& T))2)8.*&2
"./.*&*GGU:F< 7 :FED
P. U4/B.&/8D :;;F D M2)/./8*(%N4%/*% &/&2A(.(*&/0 ,-)+&-A)’.* %B,-%((.)/
)9 *=>? %/*)0./8 & ,4’&’.V% /)/3(,%*.9.* 2.,.03’-&/(9%- ,-)’%./ 9-)5 ’$%
6-&*’( )9 0)V%’-%%F 4’5%6%’ %(5#-*.1’&’ W&.22 DfD K)4-/&2 )9 X2&/’
W.)2)8A*GHFFfU1H< 7 1<:D
P. U4/B.&/8*M$%/ P. D :;;F D Y()2&’.)/ )9 ’)’&2 S>? 9-)5 0-A (%%0( )9
4’5%6%’ %(5#-*.1’&’ D K)4-/&2 )9 ")4’$Q%(’ M$./& >)-5&2 Z/.V%-(.’AU
>&’4-&2 "*.%/*%Fx,z9*))111D
"&’) #D 1CCH D I.*-),-)8&’.)/ )9 $&/+&*$./)+.F4’5%6%’ %(5#-*.1’&’fD K)4-/&2
)9 ’$% K&,&/%(% [)-%(’-A ")*.%’A*=>?@A C RH
F?s7M{A !"#@B