1. No category

‫‪Archive of SID‬‬
‫ژﻧﺘﻴﻚ ﻧﻮﻳﻦ‬
‫دوره ﭼﻬﺎرم‪ ،‬ﺷﻤﺎره ‪ ،3‬ﭘﺎﻳﻴﺰ ‪1388‬‬
‫ﺻﻔﺤﻪ ‪17-25‬‬
‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‪ -‬ﻳﻚ ‪ IGF-I‬در ﻓﻴﻞ ﻣﺎﻫﻲ‬
‫‪ Huso huso‬و ﺑﺮرﺳﻲ ﺑﻴﺎن آن در ﺑﺎﻓﺖﻫﺎي ﻣﺨﺘﻠﻒ‬
‫‬
‫
* ‬
‫ & ‪ '() !* "+, , !" #$%‬‬
‫‪.2 !3$% 45 6 !-$% ./+ 01‬‬
‫* ‪:-%-8 ./ '5- 9#:/ !/+#%‬‬
‫‪bmamiri@ut.ac.ir‬‬
‫)"‪(@@/C/ :A+B <+" @@/?/ :.+" <+‬‬
‫‬
‫ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‪-‬ﻳﻚ‪ ،IGF-I ،‬ﻧﻘﺶ ﺣﻴﺎﺗﻲ در ﻓﺮاﻳﻨﺪﻫﺎي ﻣﺨﺘﻠﻒ زﻳﺴﺘﻲ ﻣﺎﻫﻴﺎن ﺑﺮ ﻋﻬﺪه‬
‫دارد‪ .‬ﻫﺪف از اﺟﺮاي اﻳﻦ ﺗﺤﻘﻴﻖ دﺳﺘﻴﺎﺑﻲ ﺑﻪ ﺗﻮاﻟﻲ ﻛﺎﻣﻞ ژن ﻛﺪﻛﻨﻨﺪه ‪ IGF-I‬و ﺑﺮرﺳﻲ اﻟﮕﻮي‬
‫ﺑﻴﺎن آن در ﺑﺎﻓﺖﻫﺎي ﻣﺨﺘﻠﻒ ﻓﻴﻞ ﻣﺎﻫﻲ ‪ ،Huso huso‬ﺑﻪ ﻋﻨﻮان ﻳﻜﻲ از ﻣﻬﻤﺘﺮﻳﻦ ﮔﻮﻧﻪﻫﺎي ﻣﺎﻫﻴﺎن‬
‫
‬
‫ ‪،‬‬
‫‪،Huso huso‬‬
‫
‪،‬‬
‫
‪،‬‬
‫ ‬
‫ﺧﺎوﻳﺎري ﺑﻮد‪ .‬ﺑﺎ اﺳﺘﻔﺎده از روش ‪ ،RACE-PCR‬ﻗﻄﻌﻪاي ﺑﻪ ﻃﻮل ‪ 1229‬ﻧﻮﻛﻠﺌﻮﺗﻴﺪ ﺑﻪ ﻋﻨﻮان ﺗﻮاﻟﻲ‬
‫ﻛﺎﻣﻞ ‪ IGF-I‬در ﻧﻤﻮﻧﻪﻫﺎي ﻛﺒﺪي ﻓﻴﻞ ﻣﺎﻫﻲ ﺷﻨﺎﺳﺎﻳﻲ و ﺛﺒﺖ )‪ (AB512770/1‬ﮔﺮدﻳﺪ‪ .‬ﻧﺎﺣﻴﻪ‬
‫ﻛﺪﻛﻨﻨﺪه ﭘﺮوﺗﻴﻴﻦ در اﻳﻦ ﺗﻮاﻟﻲ‪ ،‬ﺷﺎﻣﻞ ‪ 483‬ﻧﻮﻛﻠﺌﻮﺗﻴﺪ اﺳﺖ‪ .‬ﭘﺮوﺗﻴﻴﻦ ﻛﺎﻫﺶ‬
‫ﻧﻴﺎﻓﺘﻪ ‪IGF-I‬‬
‫)‪ (BAH85795/1‬ﺷﺎﻣﻞ ‪ 160‬اﺳﻴﺪآﻣﻴﻨﻪ ﺑﻮده ﻛﻪ ﺑﻪ ﻃﻮر ﻛﻠﻲ از دو ﺑﺨﺶ ‪ Signal peptide‬و ﭘﻴﺶ‬
‫ﭘﺮوﺗﻴﻴﻦ ‪ (proIGF-I) IGF-I‬ﺗﺸﻜﻴﻞ ﺷﺪه اﺳﺖ‪ .‬ﻧﺎﺣﻴﻪ ﭘﻴﺶ ﭘﺮوﺗﻴﻴﻦ‪ ،‬ﺧﻮد ﺷﺎﻣﻞ ‪ 5‬ﺣﻮزه ‪D ،A ،C ،B‬‬
‫و ‪ E‬ﻣﻲﺑﺎﺷﺪ‪ .‬ﻣﻘﺎﻳﺴﻪ ﺗﻮاﻟﻲ ‪ IGF-I‬ﻓﻴﻞ ﻣﺎﻫﻲ ﺑﺎ ﺗﻮاﻟﻲ اﻳﻦ ﻓﺎﻛﺘﻮر در ﺳﺎﻳﺮ ﻣﻮﺟﻮدات‪ ،‬ﻣﺒﻴﻦ‬
‫ﺷﺒﺎﻫﺖ ﻧﺴﺒﺘﺎ ﺑﺎﻻي ‪ IGF-I‬اﻳﻦ ﻣﺎﻫﻲ ﺑﺎ دﻳﮕﺮ ﻣﻬﺮه داران ﺧﺼﻮﺻﺎ اﻧﻮاﻋﻲ از ﭘﺮﻧﺪﮔﺎن ﺑﻮد‪ .‬ﺑﺮرﺳﻲ‬
‫ﺑﻴﺎن ‪ IGF-I‬در ﺑﺎﻓﺖﻫﺎﻳﻲ ﻧﻈﻴﺮ ﻣﻌﺪه‪ ،‬آﺑﺸﺶ‪ ،‬ﻛﺒﺪ‪ ،‬ﻃﺤﺎل‪ ،‬ﻛﻠﻴﻪ‪ ،‬ﻋﻀﻠﻪ‪ ،‬ﭘﻮﺳﺖ‪ ،‬روده‪ ،‬ﻗﻠﺐ و ﻣﻐﺰ‬
‫ﺑﺎ روش ‪ Real-time PCR‬ﻧﺸﺎن داد ﻛﻪ اﮔﺮﭼﻪ اﻳﻦ ﻓﺎﻛﺘﻮر در ﺗﻤﺎﻣﻲ ﺑﺎﻓﺖﻫﺎي ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﺑﻴﺎن‬
‫ﻣﻲﺷﻮد‪ ،‬اﻣﺎ ﻣﻜﺎنﻫﺎي اﺻﻠﻲ ﺑﻴﺎن آن ﺑﻪ ﺗﺮﺗﻴﺐ ﻛﺒﺪ و روده ﻣﻲﺑﺎﺷﺪ‪ .‬ﻛﻤﺘﺮﻳﻦ ﻣﻴﺰان ﺑﻴﺎن در‬
‫ﺑﺎﻓﺖ ﻋﻀﻠﻪ اﻧﺪازهﮔﻴﺮي ﺷﺪ‪ .‬ﻧﺘﺎﻳﺞ اﻳﻦ ﺗﺤﻘﻴﻖ ﻣﻲﺗﻮاﻧﺪ ﺑﻪ ﻣﻨﻈﻮر ﺑﺮرﺳﻲ ﻓﺮاﻳﻨﺪﻫﺎي‬
‫ﻓﻴﺰﻳﻮﻟﻮژﻳﻜﻲ ﻓﻴﻞ ﻣﺎﻫﻲ از دﻳﺪﮔﺎه ﻣﻮﻟﻜﻮﻟﻲ ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﻴﺮد‪.‬‬
‫‪www.SID.ir‬‬
Archive of SID
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
A.baeri
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
R< (W) A.guldenstaedtii
* # 8*A c #7 K"$ a# (FJ428828)
#= 4 '5 75 K0 < 4 $? E $
LM "#5 IGF-I % >* .%< $"
#e "* 1# d 4"$ 2 VO*5
IGF-I * 8< 8*A 4< a5 P2#
$? '5 LM %5 K0 <
%8f I " =Y- #U < ."$
R# '5 'Q"* "$ O*5 4VO5
."#7 P5 *V*
? QV 7 $? '5 2# .%5E .9 YE a "$ N (20±140 g) *< N $? '5 8A+ g#?
'B# 7 K`a K" - "9 YER5 efD
K"$ (10ppm) MS222 b %0 hS< .""$
4J0- 4K"8 4K 4"R7 '$ LM %5 ij "D "B ."$ H& m l= 4k+ 4%<? 47 4Ta
('7 - m) 7> %5 N HE n]]
* W/\ * h? 95Y
? KE %RQ 4(Qiagen, Germany) RNAlater
;+ <*" Y$ a7 H= .BB0 7 sr]Cº 4RNA qM< "E J< S< p#
."$ "`
cDNA
#< RNAqM<
HE W]] "D '7 RNA qM< 4J D %7 K6< IGF-I "* 9 ;R# #= "R7 %5
%7$ '8*< FA (Qiagen, Germany) RNeasy
K6< M< RNA %7 %67 ."E K"<
nr] n_] J- 5VS< %W E JO ( ) & % !"#$ 2 1 .,/0 "* ,- %"+ '*
'5 4
."$ %67 '* 4Huso huso "$ :0* 9 %*85 % 4/+
5 % " 4$? @$ >? .= ;<
F- "#$ E C6D ."$ ? 2 AB
I H1 4? 8< G <
8< 2 NO*5 LM "#5 K"##7 J#7 '=
"$ 75 . "
'Q"* < P2# 4"$ 4##
K& N "S? ? %BBD 4(IGF-I) N–*1 R$
(proinsulin) *1 T? '+ < %R$ U '$ < 2 J85 %*D 75 ."$
"8 1 V= %< D A 4C 4B KD
IGF-I 9 TB KY= .(WX) %< K"$ <#$ #7
P1B K"##7 N0 "$ K"##7 P2# #=
9 '= #= 75 4LM %5 *<
"#5 8< Z 4"M JV*5 'V 8< %
Z E K D K"##7 N0 4"<
"$ [A NM E"< D < K
# "8 .8*A #7 .(W_ W\ 4W^ 4W] 4\)
2 LM E IGF-I < *
S7 (W]) Oncorhynchus mykiss 7 ` ,J+
` 1 K"$ a# (_) Cyprinus carpio *8
Ctenopharyngodon
M6= S7 2 & P E Carassius auratus
Y- (EU051323) idella
K"< %Rb (GenBank) O N (DQ777636)
.%<
LM E IGF-I TB * # *<
#
"0
1
.=Y-
4
< 4(W_ W\) Acipenser ruthenus
// / www.SID.ir
Archive of SID
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
K6< T#7 J/0 ."$ B+ W] ." xnºC
qM< RNA HEV N "B ."$ E K" JO qM< h? <V$ E JO 5V*
cDNA
iScriptTM
%7
K6<
K"$
."$ *
."$ '"R 'V DNA (Bio-Rad, Hercules, CA, USA)
\-´^´ RACE 7> O '7 * < 2# E* "B5 V V8 M1 T#7
K6<
" RNA PCR K6< O DNA - ."E (Invitrogen, Rockville, MD, USA)
.%5E +
Rapid
Amplification
cDNA
Ends
Kit HEV n/\ K6< ^´ cDNA 4" 9Y
IGF-I
AP ? EZ K "R7 K"$ qM< '7 RNA
#7 '5 IGF-I K"##7"7 O * V# QV ."$ "* SuperscriptII V8 M1 P
IGF-I3
* EZ D- "E H, 4 K"a <#$
IGF-I1 PB1 EZ K6< {" cDNA
H"+ O 8A+ QV %R1 (W J")
(Universal Amplification) UA EZ (W J")
ºC @$
RACE EZ D- %R1 K" %< * %0 V8 EZ #= ^´ sRACE %7 * 2# .E H"+ 4Real-time PCR 9/
wXºC D U ^\ K B+ n ." wX
'$ LM 75 K"$ <#$
B+ N ." xnºC b W\." \rºC 4b ^] ."
PCR96 NucleoFast®
*< (DQ201138) < R1 *
%7 K6< T#7 J/0 ."$ R< '7 * (DQ329352)
< |*
(MACHEREY-NAGEL, GmbH, Germany)
TOPO TA Cloning
%5E + 1B BLAST K6< (FJ428828)
* %7 K6< hS< K"$
%R1 4LM 1V K"$ %u6D D <
(Invitrogen, MD, USA) K"< %7$ '8*< FA
Big Dye %7
T#7 ."E H"+ Oligo v6.86 5 H EZ D-
K6< %RQ 7 ."E 7
Applied ) ABI^W^]J"
* <#$
J- O 8A+ QV 2# ? K&
VO * K`< Terminator
b W\ 4wXºC B+ @$ %0 "v7 W^n "D
.""$ * (Biosystem, CA, USA
4"$ K 2# k yzºC B+ N 4x]ºC @1 .%5E + K6< @1 EZ J/
‫ ﻃﺮاﺣﻲ آﻏﺎزﮔﺮﻫﺎ ﺑﺎ اﺳﺘﻔﺎده از ﻧﺮم اﻓﺰار‬،Huso huso ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ آﻏﺎزﮔﺮﻫﺎي ﻣﻮرد اﺳﺘﻔﺎده در ﻣﺮاﺣﻞ ﻣﺨﺘﻠﻒ ﺗﻮاﻟﻲ ﻳﺎﺑﻲ و ﺑﺮرﺳﻲ ﺑﻴﺎن‬-1 ‫ﺟﺪول‬
.‫ اﺟﺮا ﺷﺪ‬Oligo v6/86
*
٣‫و‬٢١
٣‫و‬١
٣
‫و‬٣
٢
5'-GTGCGGAGCTGAGCTTGTGGACACTCTTC-3'
5'-ACTCGTTCACGATGCCCCTGTGGTG-3'
5'-GACACGCTGCAGTTTGTGTGTGGAGA-3'
5'-GGCCACGCGTCGACTAGTACT(17T)-3'
5'-CUACUACUACUAGGCCACGCGTCGACTAGTACT-3'
5'-GCAGAAAGCCCTCCACCT-3'
5'-GCAATGGCAAATTCACCTCATCT-3'
5'-GGTAGTTCCTGTTGCCTGTGTTC-3'
IGF-I1F
IGF-I2R
IGF-I3F
AP
UAP(R)
IGF-I4R
IGF-I5R
IGF-I6R
5'-TACATCTCCAGTCGCCTCAG-3'
5'-GGCCACGCGTCGACTAGTAC-3'
IGF-I7R
Abridged Anchor (F)
// / www.SID.ir
Archive of SID
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
"* %R1 " 4IGF-I \ ´ < 2# O g <"< % . B+ N ." J#7
"R7 K"$ qM< RNA HEV n/\ 9/
cDNA
#= \´AGGAGGCCGCTGAGATGGGGAAAG^´
V8 M1 P IGF-IX EZ '5
´GTGGCCGGGAGCATCAATGATGGT´ PB1 EZ
K6< hS< .(W J") "E H"+ SuperscriptII
4" M 2# ."$ K6< V8 EZ #=
EZ #= IGF-Ix 6IGF-I 4IGF-I\ EZ
K"$ F+ B #R EZ %6 "< #0#
%7 Abridged AnchorPB1 EZ K V8
R<0 2-(Ct) {" O R1 ."$ cDNA
^\ '$ PCR LM D < \ ´ 4\´sRACE
J#7 O < 2# < K .(r) "E
b W\ ." \rºC 4b ^] ." wXºC D U
. {" O K"$ FA V FA J/0 % ."$ QV B+ N ." xnºC
"$ K T 8 {0 ± ` .9 g
."$ * 7 4< |*
}5 Elongation
factor
LM %5 #8 {Y
< 2#
Real-time PCR
h*sJV<7 5- N h &
7 LM %5 IGF-I O <
."E K6< (p<0/05) V # "#U
%R1 " 2# ."$ K6< Real-time PCR
."E H"+ (nsn "#) 'V DNA #< '7 RNA qM<
RNA
* <#$ T? 7 9/
EZ %6 N
7 67 <
EZ #= 4(W J") 5E + K6< %5 K"$ qM< RNA 67 < g 4W 'V$
P&D .%5E + K6< Real-time PCR
28S rRNA 18S & " 4 ." a * RNA HE N K"$ "* cDNA '$ µl 257
J0 7 ` g .%< K"a '+ [f
V8 PB1 EZ N , ^]] 4`*
K6< "` RNALater
Bio-) Quantitect Syber Green x2 ~M
.%< LM %5 RNA qM< 2#
*V Wn/\ P "+5 AB p *V n\ P&D B (Rad, USA
K"$ qM< RNA p> ."$ %R1
T#7 ."E < K V 4RNA K"##7 &
%$ + W/rsn/W K"0 nr] n_] q J-
'$ T#7 H& 'D ."$ iCycler Bio–Rad K`<
JR+ '+ %7 ` 7 4(%< K"a NV6 g)
U X] K wXºC P "$ J85 K B+ W\
. LM %5 RNA
xnºC b W\ ." x]ºC 4b ^] ." w\ºC D
٢٨S
١٨S
‫ و‬18S ‫ در ﺗﻤﺎﻣﻲ ﻧﻤﻮﻧﻪﻫﺎ دو ﺑﺎﻧﺪ واﺿﺢ ﻣﺘﻌﻠﻖ ﺑﻪ‬.%1 ‫ ﺑﺮ ژل آﮔﺎرز‬Huso huso ‫ اﺳﺘﺨﺮاج ﺷﺪه از ﺑﺎﻓﺖﻫﺎي ﻣﺨﺘﻠﻒ ﻓﻴﻞ ﻣﺎﻫﻲ‬RNA ‫ ﺑﺮرﺳﻲ ﻛﻴﻔﻴﺖ‬-1 ‫ﺷﻜﻞ‬
‫ آﺑﺸﺶ و ﻛﺒﺪ‬،‫ ﻗﻠﺐ‬، ‫ روده‬،‫ ﻛﻠﻴﻪ‬،‫ ﻃﺤﺎل‬،‫ ﻣﻐﺰ‬،‫ ﭘﻮﺳﺖ‬،‫ ﻣﻌﺪه‬،‫ ﻧﻤﻮﻧﻪﻫﺎ از راﺳﺖ ﺑﻪ ﭼﭗ ﺑﻪ ﺗﺮﺗﻴﺐ اﺳﺘﺨﺮاج ﺷﺪه از ﺑﺎﻓﺖﻫﺎي ﻋﻀﻠﻪ‬.‫ ﻣﺸﺎﻫﺪه ﻣﻲﺷﻮد‬28S rRNA
.‫ﻣﻲﺑﺎﺷﺪ‬
// / www.SID.ir
Archive of SID
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
'V$) "
#"< x] '5 75 '5 IGF-I * qM<
.(^
Wnnw J- 8A+ 4RACE-PCR K6< '5 IGF-I #"< "v7 * 1B
"R7 IGF-I K"##7"7 O * #= "v7
"E $ ` E "7 O N < h? qM< '5
4%w] T 4 %a 7 - .(n J")
? K"##7"7 D ."E %Rb AB\Wnxx]/W <<
.(n J") "$ K"a ` '5
nXX * h? D . "v7 Xr^ '$ * (%rx) , %R$ "v7 2 '5 IGF-I *
K"##7"7 D ? .$ Z \´-UTR "v7
'+ - 7 a 4Anser anser I 2 E"? .E + 4^´-UTR "v7 \]n * 4?
2 M< %a a sWWXW D k AATACA GATAAA *
.(%x_) Sparus aurata P< (%xX) *8S7
non-canonical
IGF-I
#= "v7 WW_\sWWxW WW^\
41 75 * '5 IGF-I %a .(n 'V$) "E <#$ polyadenylation signal
.(n J") "$ R<0 %x^
'$ '5 (BAHr\xw\/W) IGF-I 5 T7 ?
< '5 IGF-I # "< * %R$
Signal
TM 7 - ? . #"< W_]
k 7> * #e . %w] T , 1
(proIGF-I) IGF-I ? T? # "< X^ peptide
a 4I *8S7 IGF-I %a %x] xX
\ pro-IGF-I JV* .%< K"$ 'Va #"< WWx
1 7 Mus musculus %a %_]
Xx r 4nW 4Wn 4nw '$ k E D 4A 4C 4B KD
.(n J") a
h? IGF-I 75 H5 .%< K"$ 'Va #"<
J85 'V$ >* $ 'Va pro-IGF-I E KD "$"
.‫ و ﺑﺮﺧﻲ دﻳﮕﺮ از ﮔﻮﻧﻪﻫﺎي ﻣﻬﺮهداران‬Huso huso ‫ ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻣﻘﺎﻳﺴﻪ ﺷﺒﺎﻫﺖ ﻧﻮﻛﻠﺌﻮﺗﻴﺪي و اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ‬-2 ‫ﺟﺪول‬
% # "< % "v7
<< "7
E
K
Acipenseridae
ww
ww
DQ^nw^\n/W
Acipenser ruthenus
wW
wx
DQn]WW^r/W
A.gueldenstadtii
wr
wr
FJXnrrnr/W
A.baeri
xX
xX
EF\^_rrw/W
Cyprinus carpio
Cyprinidae
_X
x^
M^nxwn/WW
Oncorhynchus kisutch
Salmonidae
_w
x_
EF\_^r^x/W
Sparus aurata
Sparidae
xW
rX
DQnx\^\W/W
Gallus gallus
Phasianidae
x]
rx
EF^rn__r/W
Anser anser
Anatidae
_]
rW
AK]\]WWr/W
Mus musculus
Muridae
_^
rn
FJWw_\r^/W
Bos taurus
Bovidae
_x
x^
NM]]]_Wr/^
Homo sapience
Homonidae
// / www.SID.ir
Archive of SID
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
1
61
121
181
241
301
361
421
481
541
601
661
721
781
841
901
961
1021
1081
1141
1201
ctgtcgaaaa
gtttgctaaa
aaagtctgaa
ttttttttta
ggggatgtcc
taagatgtgc
ggctgctgca
tgtgtgtggg
gcggccacac
actggagatg
gcgccatacc
ttgcaaggag
gatttagaag
caattaaaag
ctggtgaaac
gtttaacagt
atatagccac
tgcactaagc
gcagaccttg
atggacttga
aaacacatat
tgtctgtgta
tctcgcttcc
tattgaaatg
atgacttcaa
agcaggcatt
tctttgtcct
gccggctcag
gagagaggct
cacaggggca
tactgtgcgc
gatatgccga
gtacattcga
agcaaaatcc
atgaggtgaa
gtaatataaa
gcaatgtgtt
gaagatctgg
cttttcaaag
atcaaagtag
ggacaaagag
tgacaaccgc
atgtagataa
ctctacagaa
tgacattgca
acacatttat
ccgtcttcca
gtacccatta
agactctgtg
tctatttcaa
tcgtgaacga
ctgttaagcc
aagcacaaaa
agaattccag
agagtaaaat
tttgccattg
tgagctcagt
ataggtagga
gctgtgacgt
cagcttctca
attggcacca
gttcaaatac
tccttcagg
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
atgtgaggat
aacagagtag
cttcacatct
tttgctggtc
tcagtggcac
cttgctcctg
cggagctgag
caaaccaaca
gtgctgcttc
agccaaagcc
gaaacatata
ccgcgggaac
taattataat
cagaatggtt
agtggagatg
aaatcacttt
gtgaccccat
gtgtcctgaa
gagtagcagc
agcaatggac
tttctctaaa
aagagtcagc
catcctattt
tttgtagttt
ttgtgtgatg
tgtgttctgt
cttgtggaca
ggctatgggg
cagagctgtg
tcacgctctg
tctggtaata
acaggcaaca
aacaataata
cactgtattg
caggtggagg
actaaaaatg
ggctgttggt
gttcatgtgg
aatgctgaaa
tttaagattt
tccctcttct
gcaattgaac
cgcgttggat
tgggatccat
tcttcaagtt
gcctgactca
ctcttcagtt
ccagctcgag
acctgaggcg
tacgtgccca
gccagccatc
ggaactaccg
aaaataataa
ttcatgcaaa
gctttctgct
tattgaacca
ttcatttcag
ctctgcttgt
caatggataa
tttttatttg
‫ ﻗﺒﻞ و ﺑﻌﺪ از ﻧﺎﺣﻴﻪ ﻛﺪﻛﻨﻨﺪه ﺑﻪ ﺗﺮﺗﻴﺐ‬.‫ ﻧﻮﻛﻠﺌﻮﺗﻴﺪ‬483 ‫ ﻧﺎﺣﻴﻪ ﻛﺪﻛﻨﻨﺪه ژن ﺑﺎ‬،‫ ﺣﺮوف ﺳﺎﻳﻪ دار‬.(AB512770/1) ‫ درﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﺗﻮاﻟﻲ ﻧﻮﻛﻠﺌﻮﺗﻴﺪي‬-2 ‫ﺷﻜﻞ‬
Non-canonical polyadenylation signals ‫ ﺗﻮاﻟﻲ‬AATACA ‫ و‬GATAAA ،‫ ﺗﻮاﻟﻲ ﻛﺪون ﭘﺎﻳﺎﻧﻲ‬TAG .‫ ﻧﻮﻛﻠﺌﻮﺗﻴﺪ ﻗﺮار دارد‬502 ‫ و‬244 ‫ ﺑﺎ‬UTR-´3 ‫ و‬UTR-´5
.‫ﺧﻮاﻫﺪ ﺑﻮد‬
MSSRHSVFHQWHLCDVFKFKMCSLSCTHYLLLCVLCLTQAAAAGSETLCGAELVDTLQFVCG
Signal peptide
B
ERGFYFNKPTGYGASSRRPHHRGIVNECCFQSCDLRRLEMYCAPVKPAKASRSVRAQRNTDM
B
C
A
PKAQKKHISGNSQPSCKEVHSKNSSRGNTGNRNYRI
D
E
E
43 ‫ ﺑﺎ‬signal peptide ‫( ﺷﺎﻣﻞ دو ﺑﺨﺶ اﺻﻠﻲ‬GenBank: 1/85795BAH ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ )ﻛﺪ دﺳﺘﺮﺳﻲ در‬IGF-I ‫ ﺗﻮاﻟﻲ اﺳﻴﺪآﻣﻴﻨﻪ ﭘﺮوﺗﻴﻴﻦ ﻛﺎﻫﺶ ﻧﻴﺎﻓﺘﻪ‬-3 ‫ﺷﻜﻞ‬
.‫ اﺳﻴﺪ آﻣﻴﻨﻪ‬117 ‫ ﺑﺎ‬E ‫ و‬D ،A ،C ،B ‫ ﺣﻮزه ﻣﺨﺘﻠﻒ‬5 ‫اﺳﻴﺪ آﻣﻴﻨﻪ و‬
."$ UE !%< K"$ T&#< '+ B
IGF-I ."$ a .(X 'V$) %< .6 1 %5 N#V K V8 M1 FB0 ^^ "D K h? _] "D 4"R7
%5 [A< 8 2# 4Real-time PCR
7 4l= h? .(X 'V$) "$ E K" l= 4k+ 4m 4K 4K"8 4Ta 4J0- 47 4"R7 '$ LM
k+ Ta 47 4J0- %5 k ."$
*< N $? '5 8A+ \ l= %<?
B # a < .(X 'V$) "E R<0
T&#< O R1 [A< .%5E + K6<
, #8 - K "R7 O J#7 O #= 4Elongation factor
.(p<]/]\) 5 <
."E J8 ( K"##7 KR*7 #= ) l= IGF-I < %5 7 a g
// / www.SID.ir
‫‪Archive of SID‬‬
‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‪ -‬ﻳﻚ ‪ IGF-I‬در ﻓﻴﻞ ﻣﺎﻫﻲ ‪...‬‬
‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‪ ،‬ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‪ ،‬ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬
‫‪80‬‬
‫‪70‬‬
‫‪60‬‬
‫‪40‬‬
‫‪30‬‬
‫ ‬
‫‪50‬‬
‫‪20‬‬
‫‪10‬‬
‫‪0‬‬
‫‬
‫‬
‫‬
‫‬
‫‬
‫
‬
‫‬
‫
‬
‫‬
‫‬
‫!"! ‬
‫ﺷﻜﻞ ‪ -4‬ﺷﺪت ﺑﻴﺎن ﻧﺴﺒﻲ ‪ IGF-I‬در ﻓﻴﻞ ﻣﺎﻫﻲ ‪ ،Huso huso‬ﻳﻚ ﺳﺎﻟﻪ ﭘﺮورﺷﻲ )‪ (n=5‬ﻣﺘﻌﺎدل ﺳﺎزي دادهﻫﺎ ﺑﺮ اﺳﺎس ﻣﻴﺰان ﺑﻴﺎن ‪ ،Elongation factor‬اﺟﺮا ﮔﺮدﻳﺪ‪.‬‬
‫دادهﻫﺎ ﺑﻪ ﺻﻮرت ﻣﻴﺎﻧﮕﻴﻦ ﺗﻐﻴﻴﺮ ﺑﻪ ﻧﺴﺒﺖ ﺷﺪت ﺑﻴﺎن در ﻋﻀﻠﻪ ﺑﻴﺎن ﮔﺮدﻳﺪ‪ .‬٭‪ ،‬ﻣﻘﺎدﻳﺮ ﺑﻪ ﻃﻮر ﻣﻌﻨﻲداري ﺑﺎ ﻣﻴﺰان ﺑﻴﺎن در ﺑﺎﻓﺖ ﻋﻀﻠﻪ ﻣﺘﻔﺎوت اﺳﺖ )‪.(p<0.05‬‬
‫‬
‫ ‪ IGF-I K"##7"7 O '7 * * 8*A‬‬
‫‪ E V #= '5‬‬
‫ ‪# IGF-I * # T? .%5E + <#$‬‬
‫ < ‪4R< '$ ` E‬‬
‫<* < ‪H"7 c 7 K"$ <#$‬‬
‫‪1B 4 ? 8< % k<# %+‬‬
‫ ‪."" 4 E #= '5‬‬
‫* ‪7 "v7 Wnnw '$ 7 - '5 IGF-I‬‬
‫ ‪+ ? K"##7"7 D "v7 Xr^ "8‬‬
‫‪x] '$ '5 IGF-I ? J85 'V$ .%$‬‬
‫<"‪#"< "v7 * 1B ."$ #‬‬
‫‪K"# a E < '5 IGF-I‬‬
‫‪'5 4O5 2 UE ."$ '+ %a‬‬
‫ ‪h# .6 Huso h# F8‬‬
‫‪Acipenser‬‬
‫
" ‪E O5 %a ` " .‬‬
‫>‪#e ."$ .6 h# E + PZ = 7‬‬
‫‪'95 PZ = 4R1 %R$ K"# a ` 1B‬‬
‫‪www.SID.ir‬‬
‫‪ IGF-I LM * 4 1 O5‬‬
‫‪P TB J " 7 "$ LM $‬‬
‫ ‪ k* ."$ LM IGF-I 1V RB‬‬
‫‪E"? '5 IGF-I a %R$ 42 {Y
4V‬‬
‫ ?‪*8 S7 2 M< 1B "1‬‬
‫‪ O5 %8+ K"# p "D 7 "$ K"a‬‬
‫ ‪ .%< "# K‬‬
‫‪ %+ E 1 K M< s5lZ‬‬
‫ ?"‪." *= M< 1B "1? E‬‬
‫‪ O IGF-I €/
T? a #U‬‬
‫‪%< 5E + < GH "$‬‬
‫)‪.(W‬‬
‫ ‪ '5 %5 IGF-I [A< FB0‬‬
‫‪4%5 .%5E + T&#< $? *< N‬‬
‫<‪%*, " ."$ K"a IGF-I LM [A‬‬
‫ ‪ VO*5 LM "#5 IGF-I 0 TB‬‬
‫‪ IGF-I 9/
4IGF LM TB ."$ '5‬‬
‫‪4*< P1B 2 VO*5 EE "#5‬‬
‫
‪ // /‬‬
‫‬
Archive of SID
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
.9 'Q"* < a5 P2# 4# 4"$
? QV 7 #77 %" r 4^ 4W) %< K"< .Rb LM E 47?
`<S< 7 P5 % 4YE a 7
.(W\
H= .BB0 7 `a 'D .E
TB ` 7 "$ E K" "R7 IGF-I a
S< IFAPA, El Toruno <*" Y$ a7
2 . "
IGF-I < "* "R7 0
7 FB0 4Carlos Infante BB0 %E K6<
e M< < 2 '5 "<
."E
1- Din S Y, Hurvitz A, Goldberg D, Jackson K, LevaviSivan B, Degani G (2008) Cloning of Russian sturgeon,
Acipenser gueldenstaedtii growth hormone and insulinlike growth factor I and their expression in male and
female fish during the first period of growth. Journal of
endocrinology investment. 31:201-210.
2- Duan C, Duguay S, Plisetskaya E (1993) Insulin-like
growth factor I (IGF-I) mRNA expression in coho salmon,
Oncorhynchus kisutch: tissue distribution and effects of
growth hormone/prolactin family proteins. Fish
physiology and biochemistry. 11:371-379.
3- Funes V, Asensio E, Ponce M, Infante C, Canavate JP,
Manchado M (2006) Insulin-like growth factor I and II in
the sole, Solea senegalensis: cDNA cloning and
quantitation of gene expression tissues and during larval
development.
General
and
comparative
endocrinology.149:166-172.
Perca
Sola
{< 4(x n) Oncorhynchus kisutch 7 *`#< Na67 4(Wn WW) *8S7 4(X) fluviatilis
"R7 4(r) Pagrus auriga + P< (^) senegalensis
` "$ g ."$ IGF-I 9 '0 #= 4M<s5lZ IGF-I VO*5 "#5 %R$
8A+ 2 u UE ."$ M< 4 '5
. "
B+ a < "# # 4m 4"R7 2 %5 IGF-I < 4< ."$ 7 a 7 5S 4l= 4K
4* .(W) %< .6 d %5 7 *D 4"$ E "R7 K ."$ a
IGF-I ."$ a 5S 7 %5 * * * IGF-I 7 ." a
4- Jentoft S, Aastveit AH, Andersen Ø (2004) Molecular
cloning and expression of insulin-like growth factor-I
(IGF-I) in Eurasian perch, Perca fluviatilis: lack of
responsiveness to growth hormone treatment. Fish
physiology and biochemistry. 30:67-76.
#U .(W) "$ E K" k+ l= %5 k
5- Kagawa H, Gen K, Okuzawa K, Tanaka H (2003)
Effects of luteinizing hormone and follicle-stimulating
hormone and insulin-like growth factor-I on aromatase
activity and P450 aromatase gene expression in the
ovarian follicles of red seabream, Pagrus major. Biology
of reproduction. 68: 1562-1568
6- Liang YH, Cheng CH, Chan KM (1996) Insulin-like
growth factor-I Ea2 is the predominantly expressed form
of IGF in common carp, Cyprinus carpio. Molecular
marine biology and biotechnology. 5: 145-152.
.(w n 4W) "$ d D p $
7- Pierce AL, Dickey JT, Larsen DA, Fukada H, Swanson
P, Dickhoff WW (2004) A quantitive real-time RT-PCR
assay for salmon IGF-I mRNA, and its application in the
study of GH regulation of IGF-I gene expression in
primary culture of salmon hepatocytes. General and
comparative endocrinology. 135: 401-411.
< {Y
E .6 b " 5Y
.D 4>m %8f 4 K" 2 E
"v7 * 7 %$ 9Y
- FB0 * 7 4 '5 IGF-I #"<
< IGF-I * '+ %R$ 4"E <#$
TB K"# a " 7 E"? 9/
K
LM 75 1V "D P
4*1V '5 IGF-I %*85 "* 9 '0 ."$
. "R7 %5
#= '5 IGF LM <#$ 4% TB < E .$ #a? 'Q "* "$ 2 1 LM "#5
// / www.SID.ir
Archive of SID
... ‫ در ﻓﻴﻞ ﻣﺎﻫﻲ‬IGF-I ‫ ﻳﻚ‬-‫ﺗﻮاﻟﻲﻳﺎﺑﻲ ﻓﺎﻛﺘﻮر رﺷﺪ ﺷﺒﻪ اﻧﺴﻮﻟﻴﻦ‬
‫ ﺣﻤﻴﺪ ﻓﺮﺣﻤﻨﺪ‬،‫ ﺑﺎﻗﺮ ﻣﺠﺎزياﻣﻴﺮي‬،‫ﻓﺎﻃﻤﻪ ﭘﻴﻜﺎنﺣﻴﺮﺗﻲ‬
8- Ponce M, Infante C, Funes V, Manchado M (2008)
Molecular characterization and gene expression analysis of
insulin-like growth factors I and II in the redbanded
seabream, Pagrus auriga: transcriptional regulation by
growth hormone. Comparative biochemistry and
physiology. 150: 418-426.
9- Reinecke M, Schmid A, Ermatinger R, Loffing-Cueni D
(1997) Insulin-like growth factor I in the teleost
Oreochromis mossambicus, the tilapia: Gene sequence,
tissues
expression,
and
cellular
localization.
Endocrinology. 138: 3613-3619.
10- Shamblott MJ, Chen TT (1992) Identification second
insulin-like growth factor in a fish species. Proceeding of
natural academic science. USA, 89: 8913-8917.
11- Tse MC, Vong QP, Cheng CH, Chan KM (2002) PCRcloning and gene expression studies in common carp,
Cyprinus carpio insulin-like growth factor-I. Biochemistry
and Biophysics. 1575: 63-74.
12- Vong QP, Chan KM, Cheng CH (2003) Quantification
of common carp, Cyprinus carpio IGF-I and IGF-II mRNA
by real-time PCR: differential regulation of expression by
GH. Journal of endocrinology. 178: 513-521.
13- Weber GM, Sullivan CV (2001) In vitro hormone
induction of final oocyte maturation in striped bass,
Morone saxatilis follicles is inhibited by blockers of
phyosphatidylinositol 3-kinase activity. Comparative
biochemistry and physiology. 129: 467-473.
14- Wood WA, Duan C, Bern HA (2005) Insulin-like
growth factor signaling in fish. International review of
cytology.243: 215-285.
15- Wuertz S, Gessner J, Kirschbaum F, Kloas W (2007)
Expression of IGF-I and IGF-I receptor in male and female
sterlet, Acipenser ruthenus; Evidence for an important role
in gonad maturation. Comparative biochemistry and
physiology.147: 223-230.
16- Wuertz S, Nitsche A, Jastroch M, Gessner J.,
Klingenspor M, Kirschbaum F, Kloas W (2007) The role
of the IGF-I system for vitellogenesis in maturing female
sterlet, Acipenser ruthenus Linnaeus, 1758. General and
comparative endocrinology. 150: 140–150.
// / www.SID.ir