٥
ﺲﻜﺗﻻ زا هﺪﺷاﺪﺟ زﺎﺌﺗوﺮﭘ رﺎﻬﭼ صاﻮﺧ ﻦﻴﻴﻌﺗ و يزﺎﺳاﺪﺟ
نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﺮﻴﺠﻧا ﺖﺧرد
(Ficus Carica)
ﻲﺒﺟر ﺪﻴﻣا ﺮﺘﻛد -1
ﻪﺘﺳراو ﺎﺿﺮﻟاﺪﺒﻋ ﺮﺘﻛد -2
ﻲﻜﻏﺎﺑ يﺎﻧاد داﻮﺟ -3
نﺎﻴﺗاﺮﺑ ﻲﻠﻋ -3
يﺮﻴﮕﻧﺎﻬﺟ ﻼﻴﻟ 3
هﺪﻴﻜﭼ
فﺪﻫ و ﻪﻨﻴﻣز هﺪﺷ شراﺰﮔ يدﺪﻌﺘﻣ تﻻﺎﻘﻣ رد ﺮﻴﺠﻧا ﺲﻜﺗﻻ يزﺎﺌﺗوﺮﭘ ﺖﻴﻟﺎﻌﻓ :
ﺖﺳا . فﺪﻫ ﺎﺑ ﺮﺿﺎﺣ ﻪﻌﻟﺎﻄﻣ
ﻪﺑاﺮﻴﺷ يﺎﻫزﺎﺌﺗوﺮﭘ ﻪﺴﻳﺎﻘﻣ
(Latex)
ﻪﻧﻮﮔ ﺮﻳز ﺲﻜﺗﻻ ﺎﺑ نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﺮﻴﺠﻧا ﺖﺧرد ﺎﻜﻳرﺎﻛ ﻪﻧﻮﮔ زا ﻒﻠﺘﺨﻣ يﺎﻫ
(Carica)
ﺪﺷ مﺎﺠﻧا .
ﻲﺳرﺮﺑ شور ﻦﻳا رد :
ﻪﻌﻟﺎﻄﻣ زﺎﺌﺗوﺮﭘ رﺎﻬﭼ ،ﻲﺑﺮﺠﺗ )
ﻦﻴﺴﻴﻓ ( ﻚﻤﻧ ﻚﻤﻛ ﻪﺑ يزﺎﺳاﺪﺟ يﺎﻬﺷور ﻚﻤﻛ ﻪﺑ
(Salt Fractionation)
ﻣوﺮﻛ و ﻮﻧﻮﻣ نﻮﺘﺳ يور ﺮﺑ ﻲﻓاﺮﮔﻮﺗﺎ )S
نﻮﻴﺗﺎﻛ ﺾﻳﻮﻌﺗ ﻦﻳزر ﻲﻋﻮﻧ (
جاﺮﺨﺘﺳا ﻪﺑاﺮﻴﺷ زا
ﺪﻧﺪﻳدﺮﮔ . ﺖﻴﻤﻛ هزاﺪﻧا رﺎﺑ ﻪﺳ ﻞﻗاﺪﺣ ﻪﻧﻮﻤﻧ ﺮﻫ ياﺮﺑ ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ ،ﺖﻈﻠﻏ ،ﺖﻴﻟﺎﻌﻓ نﻮﭼ ﻲﻳﺎﻫ ﺪﻧﺪﺷ يﺮﻴﮔ
.
يرﺎﻣآ نﻮﻣزآ زا هدﺎﻔﺘﺳا ﺎﺑ ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ و ﺖﻴﻟﺎﻌﻓ ﺮﻈﻧ زا ﺎﻬﻫوﺮﮔ ﻦﻴﺑ ﻪﺴﻳﺎﻘﻣ
ANOVA
رﻮﻣ ﻞﻴﻠﺤﺗ د
ﺪﻨﺘﻓﺮﮔ راﺮﻗ .
05 / 0
P<
ﻲﻨﻌﻣ ﺢﻄﺳ ناﻮﻨﻋ ﻪﺑ ﺪﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ رد يراد
.
ﻪﺘﻓﺎﻳ ﺎﻫ زﺎﺌﺗوﺮﭘ ﻦﻳا : ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ ﺮﻴﻈﻧ ﻲﺻاﻮﺧ رد ﺎﻫ
و ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ
، ﺪﻧداد نﺎﺸﻧ ﻲﻬﺑﺎﺸﻣ ﺖﻴﻟﺎﻌﻓ .
بﻮﺳر ﻚﻤﻛ ﻪﺑ ﺐﻴﺗﺮﺗ ﻪﺑ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ و ﺖﻴﻟﺎﻌﻓ شور و ﻦﻴﺋزﺎﻛ ﻲﻫد
SDS-Page
ﻦﻳا ﻦﻴﻨﭽﻤﻫ ؛ﺪﻧﺪﺷ ﻦﻴﻴﻌﺗ
ﻢﻳﺰﻧآ ﺾﻳﻮﻌﺗ نﻮﺘﺳ يور ﺮﺑ يرﺎﮔﺪﻧﺎﻣ ﺮﻈﻧ زا ﻲﺗوﺎﻔﺘﻣ رﺎﺘﻓر ﺎﻫ توﺎﻔﺗ ﻞﻴﻟد ﻪﺑ ًﻻﺎﻤﺘﺣا ﻪﻛ ﺪﻧداد نﺎﺸﻧ ﻲﻧﻮﻴﺗﺎﻛ هﺪﻨﻨﻛ
ﻲﻣ ﺎﻬﻧآ يﺪﻴﺘﭙﭘ ﻪﺸﻘﻧ و ﻦﻳدﺎﻴﻨﺑ رﺎﺘﺧﺎﺳ رد ﻲﻛﺪﻧا ﺪﺷﺎﺑ
. ﺐﻴﺗﺮﺗ ﻪﺑ ﻪﺑاﺮﻴﺷ ﻲﻤﻳﺰﻧآ ﺖﻴﻟﺎﻌﻓ و ﻦﻴﺌﺗوﺮﭘ ناﺰﻴﻣ
mg/ml
110 و
IU/ml
242 دﻮﺑ . ﺖﻴﻟﺎﻌﻓ ياراد ﻪﻛ ﻲﻳاﺰﺟا نﻮﻴﺴﻛاﺮﻓ رد ﺪﻧدﻮﺑ ﻲﻤﻳﺰﻧآ
يﺎﻫ
III-VI
ﺪﻧﺪﺷ ﺖﻓﺎﻳ .
ﻦﻴﺴﻴﻓ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ يﺎﻫ
III-VI
دوﺪﺣ 5 KDa
/ 26 هدز ﻦﻴﻤﺨﺗ ﺪﺷ
. لﻮﻠﺤﻣ 1 % ﻦﻴﺌﺗوﺮﭘ رد دﻮﺟﻮﻣ يﺎﻫ
نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
III-VI
لﻮﻠﺤﻣ رد 05M
/ 0
NH4CO3
ﻲﻟﻮﻣ بﺬﺟ ﺐﻳﺮﺿ ياراد
(ε280nm)
دوﺪﺣ رد ﺐﻴﺗﺮﺗ ﻪﺑ
22
، 5 / 21
، 21 و 9 / 20 ﺪﻧدﻮﺑ . ﻴﻟﺎﻌﻓ ﻦﻴﺴﻴﻓ ﻚﻴﺗﺎﻤﻳﺰﻧآ ﺖ ﺎﻫ
) نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
(III-VI
ياﺮﺘﺴﺑﻮﺳ زا هدﺎﻔﺘﺳا ﺎﺑ
ﺐﻴﺗﺮﺗ ﻪﺑ ﻦﻴﺋزﺎﻛ 6
/ 61 ، 7 / 62 ، 4 / 63 و 2 / 67
(Unit/mol)
ﺪﺷ ﻦﻴﻴﻌﺗ .
ﻪﺠﻴﺘﻧ يﺮﻴﮔ ﻪﺘﻓﺎﻳ سﺎﺳا ﺮﺑ:
،ﻪﻌﻟﺎﻄﻣ ﻦﻳا يﺎﻫ سﻮﻜﻴﻓ ﻒﻠﺘﺨﻣ ﻪﻧﻮﮔﺮﻳز رﺎﻬﭼ ﻦﻴﺑ رد نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﺮﻴﺠﻧا ﻪﺑاﺮﻴﺷ
ﺖﻴﻟﺎﻌﻓ و داﺪﻌﺗ ﺮﻈﻧ زا ﺎﻜﻳرﺎﻛ ﻪﻧﻮﮔﺮﻳز ﻪﺑ رﺎﻴﺴﺑ ﻦﻴﺴﻴﻓ يزﺎﺌﺗوﺮﭘ
Calimyrna
ﺪﺷر ﺎﻜﻳﺮﻣآ يﺎﻴﻧﺮﻔﻴﻟﺎﻛ رد ﻪﻛ
ﻲﻣ دراد ﺖﻫﺎﺒﺷ ،ﺪﻨﻛ .
هژاو يﺪﻴﻠﻛ يﺎﻫ ﺺﻟﺎﺧ ؛ﻦﻴﺴﻴﻓ ؛ﺮﻴﺠﻧا ﺲﻜﺗﻻ:
ﻲﻤﻳﺰﻧآ ﺖﻴﻟﺎﻌﻓ ؛يزﺎﺳ
ﺪﻨﺟﺮﻴﺑ ﻲﻜﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ﻲﻤﻠﻋ ﻪﻠﺠﻣ )
هرود 11 هرﺎﻤﺷ ، 3 لﺎﺳ ، 1383 (
هوﺮﮔ رﺎﻳدﺎﺘﺳا ؛ﻲﺘﻌﻨﺻ يزﺎﺳوراد ياﺮﺘﻛد ؛لوﺆﺴﻣ هﺪﻨﺴﻳﻮﻧ1
ﺪﻬﺸﻣ ﻲﻜﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ،يزﺎﺳوراد هﺪﻜﺸﻧاد ،ﻲﻳوراد ﻲﻤﻴﺷ ﻲﺷزﻮﻣآ
سردآ : ﺪﻬﺸﻣ - ﺪﻬﺸﻣ ﻲﻜﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد -
يزﺎﺳوراد هﺪﻜﺸﻧاد
ﻦﻔﻠﺗ : 9 - 8823255 - 0511 ﺑﺎﻤﻧ ﺮ : 8823251 - 0511 ﻲﻜﻴﻧوﺮﺘﻜﻟا ﺖﺴﭘ :
omidrajabi@yahoo.com
ﻋﻮﺑ هﺪﻜﺸﻫوﮋﭘ يژﻮﻟﻮﻧﻮﻤﻳا تﺎﻘﻴﻘﺤﺗ ﺰﻛﺮﻣ ؛يژﻮﻟﻮﻧﻮﻤﻳا ياﺮﺘﻛد2
ﺪﻬﺸﻣ ﻲﻜﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد رﺎﻴﺸﻧاد ؛ﻲﻠ
ﺪﻬﺸﻣ ﻲﻜﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ؛يزﺎﺳوراد هﺪﻜﺸﻧاد ؛يزﺎﺳوراد يﻮﺠﺸﻧاد3
٦ ﻪﻣﺪﻘﻣ سﻮﻜﻴﻓ ﺲﻨﺟ (Ficus)
ادوﺪﺣ 200 ﻮﻧ زا ﻒﻠﺘﺨﻣ ع
ﻲﻣ ﺮﺑ رد ار ﻪﻟﺪﺘﻌﻣ و هرﺎﺣ ﻖﻃﺎﻨﻣ نﺎﺘﺧرد زا ﻲﻜﻳ و دﺮﻴﮔ
ﻪﺳارﻮﻣ هداﻮﻧﺎﺧ يﺎﻬﺴﻨﺟ ﻦﻳﺮﺘﮔرﺰﺑ (Moraceae)
ﺖﺳا . رد
ﺎﻜﻳرﺎﻛ عﻮﻧ ،سﻮﻜﻴﻓ ﻢﻬﻣ عاﻮﻧا زا ﻲﻜﻳ ناﺮﻳا (Carica)
ﻲﻣ ﻲﻣ ﺪﻴﻟﻮﺗ ﺮﻴﺠﻧا ﻪﻛ ﺪﺷﺎﺑ ﺪﻨﻛ
) 1 .(
ﻪﻛ ﺮﻴﺠﻧا ﺖﺧرد ﻪﺑاﺮﻴﺷ
ﻪﺑاﺮﻴﺷ ﺢﺷﺮﺗ ﻲﻋﻮﻧ هﻮﻴﻣ و گﺮﺑ زا يا
ﺎﻫ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ ي
(Ficus carica) ﻲﻣ
،ﺪﺷﺎﺑ تاﺮﺛا ﺮﻴﻈﻧ ﻲﻧﺎﻣرد تاﺮﺛا
ﻚﻴﻤﻴﺴﻴﻠﮔﻮﭙﻴﻫ )
2 ( وﺮﺘﺴﻠﻛﻮﭙﻴﻫ ﻟ
ﻚﻴﻤ ) 3 ( بﻮﻛﺮﺳ هﺪﻨﻨﻛ
نﺎﻃﺮﺳ ) 4 ( ﻚﻴﻣﺪﻳﺮﻴﺴﻴﻠﮔ يﺮﺗﻮﭙﻴﻫ )
5 ( مﺮﻛ ﺪﺿ و )
6 ( ؛دراد
ﻢﻳﺰﻧآ ﻞﻴﻟد ﻪﺑ ﻦﻴﻨﭽﻤﻫ ،ﻪﻨﻴﻣآ يﺎﻫﺪﻴﺳا ،ﻚﻴﺘﻴﻟﻮﺌﺗوﺮﭘ يﺎﻫ
يﺮﺗ ،ﺎﻫﺪﻨﻗ ،ﻲﻧﺪﻌﻣ داﻮﻣ ﻦﭘﺮﺗ
ﺎﻫﺪﻴﺳا و ﺎﻫ نآ رد دﻮﺟﻮﻣ ﻲﻟآ ي
ﺖﺳا هدﻮﺑ نﺎﻘﻘﺤﻣ ﻪﺟﻮﺗ درﻮﻣ )
8،7 .(
ﺖﺧرد ﻪﺑاﺮﻴﺷ ﻲﺳرﺮﺑ
ﻚﻤﻛ ﻪﺑ ﺮﻴﺠﻧا SDS-Page
ﻦﻴﺌﺗوﺮﭘ دﻮﺟو ﺑ يﺎﻫ
ﺎ مﺮﺟ
ﻲﻟﻮﻜﻟﻮﻣ 25
و 48 ﺖﺒﺴﻧ ﻪﺑ و ﺮﺘﺸﻴﺑ ﺖﺒﺴﻧ ﻪﺑ نﻮﺘﻟادﻮﻠﻴﻛ
ﻦﻴﺌﺗوﺮﭘ ﺮﺘﻤﻛ زا ﺮﺘﻤﻛ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ ﺎﺑ ﻲﻳﺎﻫ
10 زا ﺮﺘﺸﻴﺑ و
100 ﺖﺳا هداد نﺎﺸﻧ ار )
9 .(
ﻦﻴﺴﻴﻓ»
ﺑ «
ﻢﻳﺰﻧآ ﻪ ﻪﺑاﺮﻴﺷ لﺎﻌﻓ ﻚﻴﺘﻴﻟﻮﺌﺗوﺮﭘ يﺎﻫ
ﻲﻣ ﻪﺘﻔﮔ سﻮﻜﻴﻓ ﺲﻨﺟ يﺎﻬﺘﺧرد دﻮﺷ
. هرﺎﺑرد دﻮﺟﻮﻣ تﺎﻋﻼﻃا
ﻲﻣ نﺎﺸﻧ نآ ﻪﻨﻴﻣز رد ﻪﻛ ﺪﻫد
،اﺮﺘﺴﺑﻮﺳ ندﻮﺑ ﻲﺻﺎﺼﺘﺧا يﺎﻫ
ﺶﻨﻛاو ،يزاﺮﺘﺳا ﺖﻴﻟﺎﻌﻓ لﺎﻌﻓ و نﻮﻴﺳاﺪﻴﺘﭙﭘ ﺲﻧاﺮﺗ يﺎﻫ
نﺪﺷ
ﻦﻳﺎﭘﺎﭘ ﻪﺑ ﻪﺑﺎﺸﻣ رﺎﻴﺴﺑ صاﻮﺧ ،هﺪﻨﻫﺎﻛ ﻞﻣاﻮﻋ ﺎﺑ (Papain)
) ﺘﻴﺳ مﺎﻧ ﺎﻳ ﻲﺘﺧرد هﺰﺑﺮﺧ هﺎﻴﮔ زا ﻞﺻﺎﺣ ﻢﻬﻣ زﺎﺌﺗوﺮﭘ ﻦﻴﺌ
(Carica Papaja ﻲﻣ نﺎﺸﻧ دﻮﺧ زا
ﺪﻫد ) 10 .(
زا يرﺎﻴﺴﺑ رد
ﺎﻫزاﺪﻴﺘﭙﭘ وﺪﻧآ ﻞﻳرﺪﻴﻔﻟﻮﺳ عاﻮﻧا زا يداﺪﻌﺗ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ عاﻮﻧا ﺖﺳا هﺪﺷ شراﺰﮔ )
11 .(
ﻲﻣ ﺮﻴﺠﻧا ﻒﻠﺘﺨﻣ يﺎﻬﻫوﺮﮔ ﺮﻳز ﻦﻴﺑ زا هﺎﻴﺳ ﺮﻴﺠﻧا ﻪﺑ ناﻮﺗ
ﺎﻳ Black Mission
، Stamforal
، و King Calimyrna
ﻲﻣ ﻮﻤﻧ و ﺪﺷر نﺎﻬﺟ ﻒﻠﺘﺨﻣ طﺎﻘﻧ رد ﻪﻛ دﺮﻛ هرﺎﺷا ﺪﻨﻨﻛ
. مزﻻ
هرﺎﺣ يﺎﻬﻠﮕﻨﺟ ﻲﺸﺣو ﺮﻴﺠﻧا ﻚﺸﺧ ﻪﺑاﺮﻴﺷ ﻪﻛ ﺖﺳا ﺮﻛذ ﻪﺑ ﻲﺑﻮﻨﺟ يﺎﻜﻳﺮﻣآ هرﺎﻗ (Ficus Glabrata)
مﺎﺧ ﻦﻴﺴﻴﻓ مﺎﻧ ﺎﺑ (Crude Ficin)
ﻲﻣ ﺪﺘﺳ و داد رازﺎﺑ رد دﻮﺷ
. هدﺎﻣ ﻦﻳا زا
جاﺮﺨﺘﺳا يﻮﻗ زﺎﺌﺗوﺮﭘ ﻦﻳﺪﻨﭼ ﺺﻟﺎﺧ و
ﻲﻣ يزﺎﺳ ﻪﻛ دﻮﺷ
ﻲﻣ ﻪﻠﻤﺟ زا ﺪﻧراد ﻊﻳﺎﻨﺻ رد ﻲﻧاواﺮﻓ دﺮﺑرﺎﻛ ﺮﻳز دراﻮﻣ ﻪﺑ ناﻮﺗ
دﺮﻛ هرﺎﺷا :
1 - ﻲﻳوراد ﻊﻳﺎﻨﺻ )
12 (
- ﻪﭽﺨﻴﻣ و ﻞﻴﮔز هﺪﻨﻨﻛ فﺮﻃ ﺮﺑ نﻮﻴﺳﻻﻮﻣﺮﻓ رد هدﺎﻔﺘﺳا
- نﻮﻴﺳﻻﻮﻣﺮﻓ رد هدﺎﻔﺘﺳا مﺮﻛ يﺎﻫ
ﻒﻴﻄﻟ ﺖﻬﺟ ﺎﻫ ندﺮﻛ
ﺖﺳﻮﭘ
2 - ﻲﻳاﺬﻏ ﻊﻳﺎﻨﺻ )
14،13 (
- د هدﺎﻔﺘﺳا ﺖﺷﻮﮔ ندﺮﻛدﺮﺗ ﺖﻬﺟ ،ﺖﺷﻮﮔ ﻊﻳﺎﻨﺻ ر
يﺎﻫ
ﺲﻴﺳﻮﺳ و سﺎﺒﻟﺎﻛ ﻪﻴﻬﺗ و ﺰﭘﺮﻳد
- يزﺎﺳﺮﻴﻨﭘ ﺖﻌﻨﺻ رد هدﺎﻔﺘﺳا
3 - ﻲﻫﺎﮕﺸﻳﺎﻣزآ و ﻲﻜﺷﺰﭘ ﻊﻳﺎﻨﺻ )
15 (
- ﺖﻴﻛ ﺖﺧﺎﺳ رد هدﺎﻔﺘﺳا ﻦﻴﻴﻌﺗ يﺎﻫ
نﻮﺧ ﻚﻧﺎﺑ رد Rh
ﺮﻴﺠﻧا ﺖﺧرد ﺲﻜﺗﻻ رد دﻮﺟﻮﻣ يﺎﻫزﺎﺌﺗوﺮﭘ عاﻮﻧا ﺖﺧﺎﻨﺷ ﻲﭘ و نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﻴﺑ ﺖﻴﻟﺎﻌﻓ ﻪﺑ ندﺮﺑ
نﻮﻨﻛﺎﺗ ﺎﻬﻧآ ﻲﻜﻳژﻮﻟﻮ
ﻲﻣ ﻞﺻﺎﺣ تﺎﻋﻼﻃا و ﺖﺳا ﻪﺘﻓﺮﮕﻧ راﺮﻗ ﻖﻴﻘﺤﺗ درﻮﻣ ﺪﻧاﻮﺗ
ﺪﻫد يرﺎﻳ ار قﻮﻓ ﻊﻳﺎﻨﺻ نﺎﻘﻘﺤﻣ .
ﻪﺑاﺮﻴﺷ يﺎﻫزﺎﺌﺗوﺮﭘ ﻪﺴﻳﺎﻘﻣ فﺪﻫ ﺎﺑ ﺮﺿﺎﺣ ﻪﻌﻟﺎﻄﻣ (Latex) ﻪﻧﻮﮔ ﺮﻳز ﺲﻜﺗﻻ ﺎﺑ نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﺮﻴﺠﻧا ﺖﺧرد
يﺎﻫ
ﻪﻧﻮﮔ زا ﻒﻠﺘﺨﻣ Carica
ﺪﺷ مﺎﺠﻧا .
ﻲﺳرﺮﺑ شور
ﺠﺗ ﻪﻌﻟﺎﻄﻣ ﻦﻳا رد
،ﻲﺑﺮ ﻪﺑ هدﺎﻔﺘﺳا درﻮﻣ ﺮﻴﺠﻧا ﻪﺑاﺮﻴﺷ
رد يزﺎﺳوراد هﺪﻜﺸﻧاد فاﺮﻃا ﺮﻴﺠﻧا نﺎﺘﺧرد زا هزﺎﺗ ترﻮﺻ ﺪﻬﺸﻣ ) ناﺮﻳا 1382 ( ﻊﻤﺟ رد ﻲﻫﺎﺗﻮﻛ تﺪﻣ ياﺮﺑ نآ زا و يروآ
يﺎﻣد 20 - ﺪﺷ يراﺪﻬﮕﻧ داﺮﮕﻴﺘﻧﺎﺳ ﻪﺟرد .
ﻲﺳودﺮﻓ هﺎﮕﺸﻧاد مﻮﻳرﺎﺑﺮﻫ ﻂﺳﻮﺗ ﺖﺧرد ﻪﻧﻮﮔ و ﺲﻨﺟ ﺪﻳدﺮﮔ ﻦﻴﻴﻌﺗ ﺪﻬﺸﻣ .
ﺳ ،ﻦﻴﺴﻴﻓ ،ﻦﻳﺎﭘﺎﭘ ،تﺎﻔﻟﻮﺳ ﻞﻴﺳدود ﻢﻳﺪ
ﺖﻛﺮﺷ زا ﺲﻳﺮﺗ ،ﺪﻴﻣآ ﻞﻳﺮﻛآ ﺲﻴﺑ ،ﺪﻴﻣآ ﻞﻳﺮﻛآ Sigma
) نﺎﻤﻟآ ( ﻲﺑ مﻮﻴﻧﻮﻣآ ،ﺪﻴﺳا ﻚﻴﺘﺳاوﺮﻠﻛ يﺮﺗ ،ﻦﻴﺋزﺎﻛ ، ،تﺎﻨﺑﺮﻛ
٧
ﺖﻛﺮﺷ زا ﺪﻳاﺮﻠﻛ ﻢﻴﺴﻠﻛ ،تﺎﻔﺴﻓ ﻢﻳﺪﺳ ،ﺪﻳاﺮﻠﻛ ﻢﻳﺪﺳ Merck
) نﺎﻤﻟآ ( ﺮﮕﻧﺎﺸﻧ و (Marker)
زا ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ دراﺪﻧﺎﺘﺳا
ﺖﻛﺮﺷ Invitrogen )
ﺎﻜﻳﺮﻣآ ( راﺪﻳﺮﺧ ﺪﺷ ي .
ﺪﻳدﺮﮔ ﻦﻴﻴﻌﺗ درﻮﻓداﺮﺑ شور ﻪﺑ دﻮﺟﻮﻣ ﻦﻴﺌﺗوﺮﭘ ناﺰﻴﻣ ) 16 .(
ناﺰﻴﻣ ﻦﻴﻴﻌﺗ رد دراﺪﻧﺎﺘﺳا ﻦﻴﺌﺗوﺮﭘ ناﻮﻨﻋ ﻪﺑ ﻦﻳﺎﭘﺎﭘ زا ﺪﺷ هدﺎﻔﺘﺳا ﻦﻴﺌﺗوﺮﭘ .
،زرﻮﻓوﺮﺘﻜﻟا ﺖﻬﺟ SDS-Page
ﺎﺑ هﺪﻨﻨﻛﺎﻴﺣا ﻂﻳاﺮﺷ رد
نژﻮﻤﻫ هﺪﻨﻨﻛاﺪﺟ لژ زا هدﺎﻔﺘﺳا 5
/ 12 % هﺪﻨﻨﻛ ﻊﻤﺟ لژ و 5
%
ﺷ هاﺮﻤﻫ ﻪﺑ ﺖﺑﺎﺛ ژﺎﺘﻟو رد مﺎﺠﻧا ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ دراﺪﻧﺎﺘﺳا ﺺﺧﺎ
ﺪﻳدﺮﮔ ) 13 .(
ﮓﻧر لژ يﺰﻴﻣآ هﺮﻘﻧ تاﺮﺘﻴﻧ شور زا هدﺎﻔﺘﺳا ﺎﺑ ﺎﻫ
ﺖﻓﺮﮔ ترﻮﺻ .
عﻮﻧ زا هدﺎﻔﺘﺳا درﻮﻣ هﺎﮕﺘﺳد Bio-Rad
) ﺎﻜﻳﺮﻣآ ( دﻮﺑ .
ﻦﻴﺴﻴﻓ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ ﻚﻤﻛ ﻪﺑ ﺎﻫ
SDS-Page ﻦﻴﻤﺨﺗ
ﺪﺷ هدز . ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ ﻢﺘﻳرﺎﮕﻟ ياﺮﺑ دراﺪﻧﺎﺘﺳا ﻲﻨﺤﻨﻣ
ﻣ رد ﺎﻫﺮﮕﻧﺎﺸﻧ ﺮﻳدﺎﻘﻣ ﻞﺑﺎﻘ
ﺪﻳدﺮﮔ ﻢﺳرRf )
17 .(
ﺎﺑ ﺰﻴﻟﺎﻳد ﻪﺴﻴﻛ ﻂﺳﻮﺗ ﺰﻴﻟﺎﻳد 10000
Cut Off=
ﺖﻛﺮﺷ
Sigma ) ﺎﻜﻳﺮﻣآ ( ﺪﺷ مﺎﺠﻧا .
،يزﺎﺳ ﺺﻟﺎﺧ و جاﺮﺨﺘﺳا رﻮﻈﻨﻣ ﻪﺑ 15ml
رد ﻪﺑاﺮﻴﺷ
25ml تﺎﻔﺴﻓ ﺮﻓﺎﺑ 7
= ﺪﺷ نﻮﻴﺴﻧﺎﭙﺳﻮﺳ pH .
نﻮﻴﺴﻧﺎﭙﺳﻮﺳ
رد ﺖﻋﺎﺳ ﻪﺳ تﺪﻣ ياﺮﺑ ﻪﻠﺻﺎﺣ يﺎﻣد
4 داﺮﮕﻴﺘﻧﺎﺳ ﻪﺟرد
ﺎﺳ ﺲﭙﺳ و طﻮﻠﺨﻣ اﺪﺟ نآ لﻮﻠﺤﻣﺎﻧ داﻮﻣ ﺎﺗ ﺪﻳدﺮﮔ ژﻮﻔﻳﺮﺘﻧ
ددﺮﮔ . ﻞﻤﻋ ،ﺪﻌﺑ ﻪﻠﺣﺮﻣ رد Salting Out
ندﺮﻛ ﻪﻓﺎﺿا ﺎﺑ
7 / 17 ﻪﺑ ﻢﻳﺪﺳ ﺪﻳﺮﻠﻛ 100ml
ﺪﻳدﺮﮔ مﺎﺠﻧا فﺎﻔﺷ لﻮﻠﺤﻣ زا .
ﺖﺷﺬﮔ زا ﺪﻌﺑ 10
يﺎﻣد رد ﺖﻋﺎﺳ 4
بﻮﺳر ،داﺮﮕﻴﺘﻧﺎﺳ ﻪﺟرد
تﺎﻔﺴﻓ ﺮﻓﺎﺑ رد هرﺎﺑود و ﺪﻳدﺮﮔ اﺪﺟ ژﻮﻔﻳﺮﺘﻧﺎﺳ ﺎﺑ )
6 = و pH
05 M / 0 و 3ml ( ﺪﺷ هﺪﻨﻛاﺮﭘ .
تﺪﻣ ﻪﺑ ﺲﭙﺳ طﻮﻠﺨﻣ ﻦﻳا
رد ﺐﺷ ﻚﻳ 4
تﺎﻔﺴﻓ ﺮﻓﺎﺑ ﻞﺑﺎﻘﻣ رد داﺮﮕﻴﺘﻧﺎﺳ ﻪﺟرد )
6 = pH
و 05M / 0 ( ﺪﻳدﺮﮔ ﺰﻴﻟﺎﻳد .
ﻪﺴﻴﻛ رد دﻮﺟﻮﻣ لﻮﻠﺤﻣﺎﻧ داﻮﻣ ،ﺰﻴﻟﺎﻳد مﺎﺠﻧا زا ﺪﻌﺑ
لﻮﻠﺤﻣ ﺶﺨﺑ و ﺪﺷ اﺪﺟ ژﻮﻔﻳﺮﺘﻧﺎﺳ ﻚﻤﻛ ﻪﺑ ﺰﻴﻟﺎﻳد )
دوﺪﺣ ﻪﻛ
5ml دﻮﺑ ( هﺎﮕﺘﺳد ﻪﺑ )FPLC
نﻮﺘﺳ ﻪﺑ ﺰﻬﺠﻣ 10]
/ 10 [Mono-S HR ﺾﻳﻮﻌﺗ عاﻮﻧا زا ،
هﺪﻨﻨﻛ ﻲﻧﻮﻴﺗﺎﻛ يﺎﻫ (
زﺎﻓ ﺎﺑ ﻪﻛ
تﺎﻔﺴﻓ ﺮﻓﺎﺑ كﺮﺤﺘﻣ )
6 = و pH 05 M
/ 0 ( هﺪﻴﺳر لدﺎﻌﺗ ﻪﺑ
ﺪﺷ ﻖﻳرﺰﺗ ،دﻮﺑ .
هﺎﮕﺘﺳد عﻮﻧ ﻪﺑ FPLC
ﻪﺘﻓر رﺎﻛ Pharmacia
) ﻪﺴﻧاﺮﻓ (
دﻮﺑ . ﻪﻛ ﺖﺳا ﺮﻛذ ﻪﺑ مزﻻ ﻞﻤﻋ مﺎﺠﻧا ياﺮﺑ ﻲﻫﺎﮕﺘﺳدFPLC
ﺮﻜﻠﻤﻋ ﺮﻈﻧ زا و ﺖﺳا ﻲﻓاﺮﮔﻮﺗﺎﻣوﺮﻛ ﻪﺑ يدﺎﻳز رﺎﻴﺴﺑ ﺖﻫﺎﺒﺷ د
هﺎﮕﺘﺳد درادHPLC
.
تﺎﻔﺴﻓ ﺮﻓﺎﺑ ﺎﺑ نﻮﺘﺳ ﺲﭙﺳ )
8 / 6 = و pH 005 M
/ 0 و
30ml ( ﻢﻳﺪﺳ ﻲﻄﺧ نﺎﻳداﺮﮔ زا هدﺎﻔﺘﺳا ﺎﺑ ﺶﻳﻮﺷ و ﻪﺘﺴﺷ
زا تﺎﻔﺴﻓ 8
/ 6 = و pH 005 M
/ 0 ﺎﺗ 2 / 6 = و pH
185 M / 0 ﺖﻓﺎﻳ ﻪﻣادا .
ﻪﻛ دﻮﺑ هﺪﺷ ﻢﻴﻈﻨﺗ يﻮﺤﻧ ﻪﺑ نﺎﻳداﺮﮔ
رﻮﺒﻋ ﺎﺑ 100ml
نﻮﺘﺳ زا لﻮﻠﺤﻣ ﻪﺑ تﺎﻔﺴﻓ ﻢﻳﺪﺳ ﺖﻈﻠﻏ
زا ﻲﻄﺧ ترﻮﺻ 005 M
/ 0 ﺎﺗ 185 M / 0 ﺪﻨﻛ ﺮﻴﻴﻐﺗ .
ﻦﻳا
نﻮﻴﺴﻛاﺮﻓ و ﺪﺷ مﺎﺠﻧا قﺎﺗا يﺎﻣد رد تﺎﻴﻠﻤﻋ ﺖﻋﺮﺳ ﺎﺑ ﺎﻫ
نﺎﻳﺮﺟ 60ml/h
ﻊﻤﺟ ﺪﻧﺪﺷ يروآ )
18 .(
نﻮﻴﺴﻛاﺮﻓ ﻪﻣادا رد ﻪﻛ ﻲﺠﻨﺳرﺎﻴﻋ يﺎﻬﺷور ﻚﻤﻛ ﻪﺑ ﺎﻫ
،ﺪﺷ ﺪﻫاﻮﺧ هداد حﺮﺷ ﺪﻧﺪﺷ ﻲﺳرﺮﺑ
. ﻪﻟﻮﻟ ياراد يﺎﻫ
نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
III-VI ) ﻞﻜﺷ 1 ( ﻪﻓﺎﺿا ﺎﺑ و هﺪﻳدﺮﮔ طﻮﻠﺨﻣ
ندﺮﻛ 35 رد ﺪﻳﺮﻠﻛ ﻢﻳﺪﺳ مﺮﮔ 100 ml
لﻮﻠﺤﻣ
(Salting-Out) ﺪﻧﺪﺷ هداد بﻮﺳر
. ﻦﻴﺌﺗوﺮﭘ هدﺮﻛ بﻮﺳر يﺎﻫ
تﺎﻔﺴﻓ ﺮﻓﺎﺑ رد و هﺪﺷ ژﻮﻔﻳﺮﺘﻧﺎﺳ )
8 / 6 = و pH 005 M
/ 0 (
ﺪﻧﺪﺷ هﺰﻴﻠﻴﻓﻮﻴﻟ .
ﻞﻜﺷ 1 - ﺲﻜﺗﻻ زا هﺪﺷ اﺪﺟ تﺎﺑﻮﺳر ماﺮﮔﻮﺗﺎﻣوﺮﻛ
) ﺳا ﺎﺑ ﻚﻤﻧ ﺎﺑ يزﺎﺳاﺪﺟ شور زا هدﺎﻔﺘ (
نﻮﺘﺳ يور Mono-S
و ﺖﻈﻠﻏ نﺎﻳداﺮﮔ ﺎﺑ تﺎﻔﺴﻓ ﺮﻓﺎﺑpH
٨ ﻦﻴﺴﻴﻓ ،ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ ﻦﻴﻴﻌﺗ ﺖﻬﺟ هﺰﻴﻠﻴﻓﻮﻴﻟ يﺎﻫ
يﺎﻣد رد ﺐﺷ ﻚﻳ تﺪﻣ ﻪﺑ و ﻞﺣ بآ رد هﺪﺷ 4
ﻪﺟرد
ﺪﻧﺪﺷ ﺰﻴﻟﺎﻳد بآ ﻞﺑﺎﻘﻣ رد داﺮﮕﻴﺘﻧﺎﺳ .
لﻮﻠﺤﻣ ﻦﻳا ًادﺪﺠﻣ ﺎﻫ
ﺪﻧﺪﻳدﺮﮔ هﺰﻴﻠﻴﻓﻮﻴﻟ .
ﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ لﻮﻠﺤﻣ ﻪﻴﻬﺗ ﺎﺑ ﻲﻟﻮ
(w/v) 1
% ﻪﻧﻮﻤﻧ زا ﻦﻴﺴﻴﻓ هﺪﺷ هﺰﻴﻠﻴﻓﻮﻴﻟ يﺎﻫ
ﺎﻫ
) نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
III-VI ( لﻮﻠﺤﻣ رد 05
/ 0 ﻲﺑ مﻮﻴﻧﻮﻣآ تﺎﻨﺑﺮﻛ
ﺪﻳدﺮﮔ ﻦﻴﻴﻌﺗ .
لﻮﻠﺤﻣ ﻦﻳا بﺬﺟ رد ﺎﻫ
جﻮﻣ لﻮﻃ 280 nm
ﻪﺑ
ﺮﺘﻣﻮﺘﻓوﺮﺘﻜﭙﺳا ﻚﻤﻛ (Shimatzu-Japan CPS-240A)
رد
ﺪﺷ ﻦﻴﻴﻌﺗ قﺎﺗا يﺎﻣد )
15 .(
وﺮﭘ ﺖﻴﻟﺎﻌﻓ ﻲﺳرﺮﺑ ﻢﻳﺰﻧآ ﻚﻴﺘﻴﻟﻮﺌﺗ
شور ﻚﻤﻛ ﻪﺑ ﺎﻫ
Kuntiz ﺪﺷ مﺎﺠﻧا
) 14 .(
لﻮﻠﺤﻣ 1 / 0 ﻢﻳﺪﺳ ﺮﻓﺎﺑ رﻻﻮﻣ
تﺎﻔﺴﻓ 7
= هاﺮﻤﻫ ﻪﺑ ﻦﻴﺴﻴﻓ زا ﻲﻨﻴﻌﻣ يﺎﻬﺘﻈﻠﻏ ياراد pH
NaEDTA 001M
/ 0 و 5 / 0 % ﻲﻳﺎﻬﻧ ﻢﺠﺣ ﺎﺑ ﻦﻴﺋزﺎﻛ زا
20ml تﺪﻣ ياﺮﺑ ، 20
يﺎﻣد رد ﻪﻘﻴﻗد 37
داﺮﮕﻴﺘﻧﺎﺳ ﻪﺟرد
ﺪﺷ ﻪﺑﻮﻜﻧا .
ﺶﻨﻛاو ﻪﻓﺎﺿا ﺎﺑ يﺮﺗ ندﺮﻛ
ﺪﻴﺳا ﻚﻴﺘﺳا وﺮﻠﻛ
) 5V/V % و 3ml ( بﺬﺟ ،ژﻮﻔﻳﺮﺘﻧﺎﺳ مﺎﺠﻧا زا ﺪﻌﺑ و ﺪﺷ ﻒﻗﻮﺘﻣ
رد ﻲﻳور ﻊﻳﺎﻣ جﻮﻣ لﻮﻃ
250nm ﺪﻳدﺮﮔ ﺖﺋاﺮﻗ .
ﺪﺣاو ﺮﻫ
ﻲﻣ ﻖﻠﻌﺗ ﻢﻳﺰﻧآ زا ﻲﻧاﺰﻴﻣ ﻪﺑ ﺖﻴﻟﺎﻌﻓ ﻲﺑﺬﺟ ﺮﻴﻴﻐﺗ ﺪﻧاﻮﺘﺑ ﻪﻛ دﺮﻴﮔ
هزاﺪﻧا ﻪﺑ 1
جﻮﻣ لﻮﻃ رد ﺪﺣاو 250 nm
دﺎﺠﻳا ﻲﻳور ﻊﻳﺎﻣ رد
ﺪﻨﻛ ) 19 .(
ﺻ ﺶﻳاﺰﻓا ﺖﻬﺟ ،هﺪﻣآ ﺖﺳد ﻪﺑ ﺞﻳﺎﺘﻧ ﺖﺤ
ﺖﻴﻤﻛ ياﺮﺑ ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ ،ﺖﻈﻠﻏ ،ﺖﻴﻟﺎﻌﻓ نﻮﭼ ﻲﻳﺎﻫ
هزاﺪﻧا رﺎﺑ ﻪﺳ ﻞﻗاﺪﺣ ﻪﻧﻮﻤﻧ ﺮﻫ ﺪﻧﺪﺷ يﺮﻴﮔ
. ﻦﻴﺑ ﻪﺴﻳﺎﻘﻣ
زا هدﺎﻔﺘﺳا ﺎﺑ ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ و ﺖﻴﻟﺎﻌﻓ ﺮﻈﻧ زا ﺎﻬﻫوﺮﮔ يرﺎﻣآ نﻮﻣزآ ANOVA
ﺪﻨﺘﻓﺮﮔ راﺮﻗ ﻞﻴﻠﺤﺗ درﻮﻣ .
ﻪﺘﻓﺎﻳ ﺎﻫ ﻳﺰﻧآ ﺖﻴﻟﺎﻌﻓ و ﻦﻴﺌﺗوﺮﭘ ناﺰﻴﻣ ﺐﻴﺗﺮﺗ ﻪﺑ ﻪﺑاﺮﻴﺷ ﻲﻤ
mg/ml 110
و IU/ml 242 دﻮﺑ . فﺎﻔﺷ هرﺎﺼﻋ ماﺮﮔﻮﺗﺎﻣوﺮﻛ
ﻞﻜﺷ رد ،ﺪﺷ اﺪﺟ ﺪﻳاﺮﻠﻛ ﻢﻳﺪﺳ ﻚﻤﻛ ﻪﺑ ﻪﻛ ﻪﺑاﺮﻴﺷ 1
نﺎﺸﻧ ،
ﺖﺳا هﺪﺷ هداد .
نﻮﻴﺴﻛاﺮﻓ رد ،ﺪﻧدﻮﺑ ﻲﻤﻳﺰﻧآ ﺖﻴﻟﺎﻌﻓ ياراد ﻪﻛ ﻲﻳاﺰﺟا يﺎﻫ
III-VI ﺪﻧﺪﺷ ﺖﻓﺎﻳ .
نﻮﻴﺴﻛاﺮﻓ ﻦﻳا زا ﻚﻳ ﺮﻫ زرﻮﻓوﺮﺘﻜﻟا ﺎﻫ
ﻲﺼﻟﺎﺧ ﺐﻴﻛﺮﺗ داد نﺎﺸﻧ ار
. ﻪﻟﻮﻟ ياراد ﻪﻛ ﻲﻳﺎﻫ
نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
III-VI ) ﻞﻜﺷ 1 ( ﻪﻧﺎﮔاﺪﺟ رﻮﻃ ﻪﺑ ،ﺪﻧدﻮﺑ
ﺪﻧﺪﺷ هﺰﻴﻠﻴﻓﻮﻴﻟ ﺲﭙﺳ و طﻮﻠﺨﻣ .
ﻦﻴﺴﻴﻓ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ يﺎﻫ
III-VI دوﺪﺣ 5KDa / 26 هدز ﻦﻴﻤﺨﺗ ﺪﺷ
) ﻞﻜﺷ 2 .(
لﻮﻠﺤﻣ 1
% ﻦﻴﺌﺗوﺮﭘ نﻮﻴﺴﻛاﺮﻓ رد دﻮﺟﻮﻣ يﺎﻫ يﺎﻫ
III-VI لﻮﻠﺤﻣ رد 05M
/ 0 NH4CO3 ﺮﺿ ياراد
بﺬﺟ ﺐﻳا
ﻲﻟﻮﻣ (ε280nm) دوﺪﺣ رد ﺐﻴﺗﺮﺗ ﻪﺑ
22 ، 5 / 21 ، 21 و 9 / 20
ﺪﻧدﻮﺑ . ﻦﻴﺴﻴﻓ ﻚﻴﺗﺎﻤﻳﺰﻧآ ﺖﻴﻟﺎﻌﻓ ﺎﻫ
) نﻮﻴﺴﻛاﺮﻓ يﺎﻫ
(III-VI ﺎﺑ
ﺐﻴﺗﺮﺗ ﻪﺑ ﻦﻴﺋزﺎﻛ ياﺮﺘﺴﺑﻮﺳ زا هدﺎﻔﺘﺳا 6
/ 61 ، 7 / 62 ، 4 / 63 و
2 / 67 (Unit/mol) ﺪﺷ ﻦﻴﻴﻌﺗ
) لوﺪﺟ 1 .(
ﻪﺠﻴﺘﻧ و ﺚﺤﺑ يﺮﻴﮔ
ﭼ يوﺎﺣ ﻲﻃﻮﻠﺨﻣ ﺮﻴﺠﻧا ﺖﺧرد ﻪﺑاﺮﻴﺷ ﻲﻨﻴﺌﺗوﺮﭘ ءﺰﺟ ﻦﻳﺪﻨ
ﺖﺳا . زا ﺮﺘﺸﻴﺑ ،ﻖﻴﻘﺤﺗ ﻦﻳا رد 7
ﻪﺑاﺮﻴﺷ رد ﻲﻨﻴﺌﺗوﺮﭘ ءﺰﺟ
ﺪﺷ يزﺎﺳاﺪﺟ ﻲﻓاﺮﮔﻮﺗﺎﻣوﺮﻛ شور ﻪﺑ ﺮﻴﺠﻧا .
ءﺰﺟ زا ﺮﻴﻏ ﻪﺑ I
ﺪﻧداد نﺎﺸﻧ ار ﻲﻨﻴﺌﺗوﺮﭘ ءﺰﺟ ﻚﻳ ﺎﻬﻨﺗ زرﻮﻓوﺮﺘﻜﻟا رد اﺰﺟا ﺮﻳﺎﺳ .
ياﺰﺟا وI وII ﺪﻧداﺪﻧ نﺎﺸﻧ ﻚﻴﺘﻟﻮﺌﺗوﺮﭘ ﺖﻴﻟﺎﻌﻓVII .
و ﻦﻴﻋ رد ﻦﻴﺴﻴﻓ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ رد توﺎﻔﺗ دﻮﺟ
دﻮﺟﻮﻣ يﺎﻫ
ﺎﺗاﺮﺑﻼﮔ ﻪﻧﻮﮔ رد )
1 ( ياﺰﺟا ، III-VI ﺮﻴﺠﻧا ﻪﺑاﺮﻴﺷ زا هﺪﺷ اﺪﺟ
ﺪﻧداد نﺎﺸﻧ ﻲﻧﺎﺴﻜﻳ ﻲﻟﻮﻜﻟﻮﻣ مﺮﺟ نﺎﺳاﺮﺧ ﻲﻣﻮﺑ )
5KD / 26 (
ﻲﻣ ﺮﻈﻧ ﻪﺑ ﻪﻛ ﺪﻨﺷﺎﺑ ﻦﻴﺴﻴﻓ ،ﺪﺳر
. رد ﻲﺗوﺎﻔﺗ ﻖﻴﻘﺤﺗ ﻦﻳا رد
ﻦﻴﺴﻴﻓ ﺖﻴﻟﺎﻌﻓ ﺸﻧ هﺪﻫﺎﺸﻣ ﺎﻬﻧآ ﻲﻟﻮﻣ ﻲﺷﻮﻣﺎﺧ ﺐﺋاﺮﺿ و ﺎﻫ
ﺪ
) 05 / 0 P<
.(
ﻞﻜﺷ 2 - ﻦﻴﺴﻴﻓ ﻲﻟﻮﻜﻟﻮﻣ نزو ﻦﻴﻤﺨﺗ ﻪﺑ
شور
زرﻮﻓوﺮﺘﻜﻟا SDS-Page
y = -0.1732x + 5.5429
R 2 = 0.9942
3.5
4
4.5
5
5.5
1 2 3 4 5 6 7
Relative Mobility log
(M
W)
Ficins
٩
ﻪﻌﻟﺎﻄﻣ رد لﺎﺳ رد يا
1964 سﻮﻜﻴﻓ زا ﻪﻧﻮﮔﺮﻳز ﺪﻨﭼ ،
ﺎﻜﻳرﺎﻛ
(Stanford, King, Black Mission, Calimyrna)
؛ﺪﻨﺘﻓﺮﮔ راﺮﻗ ﻪﺴﻳﺎﻘﻣ درﻮﻣ ﻢﻳﺰﻧآ يزﺎﺳاﺪﺟ ﻖﻴﻘﺤﺗ ﻦﻳا رد
يﺎﻫ
ﮔﺮﻳز ﻦﻳا ﻪﺑاﺮﻴﺷ رد دﻮﺟﻮﻣ ﻪﻧﻮ
هدﺎﻔﺘﺳا ﺎﺑ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ زا ﺎﻫ
ﻲﻓاﺮﮔﻮﺗﺎﻣوﺮﻛ نﻮﺘﺳ زا CM- Cellulose
ﺪﺷ مﺎﺠﻧا )
11 ( ،
) ﻞﻜﺷ 3 .(
ﺎﻬﻧآ ﺖﻴﻟﺎﻌﻓ هاﺮﻤﻫ ﻪﺑ هﺪﺷ اﺪﺟ ﻲﻤﻳﺰﻧآ لﺎﻌﻓ ياﺰﺟا
ﺐﺴﺣﺮﺑ Unit/mg لوﺪﺟ رد
2 ﻲﻣ ﻪﻈﺣﻼﻣ دﻮﺷ
.
ﺮﻴﺠﻧا ﻪﺑاﺮﻴﺷ يزﺎﺌﺗوﺮﭘ يﻮﮕﻟا زا هﺪﻣآ ﺖﺳد ﻪﺑ ﻪﺴﻳﺎﻘﻣ ﺎﺑ ﻪﻧﻮﮔﺮﻳز ﺎﺑ نﺎﺳاﺮﺧ ﻲﻣﻮﺑ ﻫ
لوﺪﺟ رد هﺪﺷ هرﺎﺷا يﺎ 2
و
هﻮﻴﻣ و ﺎﻬﮔﺮﺑ ﻞﻜﺷ و ﻲﮔرﺰﺑ ﺮﻈﻧ زا ﻦﻴﻨﭽﻤﻫ ﺪﻳآ ﺮﻈﻧ ﻪﺑ ،ﺎﻫ
ﻪﻧﻮﮔﺮﻳز ﻪﺑ ﻲﭙﻴﺗﻮﻧژ ﺮﻈﻧ زا ﺮﻴﺠﻧا ﻦﻳا Calimyrna
رد ﻪﻛ
ﻲﻣ ﺎﻜﻳﺮﻣآ يﺎﻴﻧﺮﻔﻴﻟﺎﻛ ﺖﻟﺎﻳا ﺖﺳا ﻪﻴﺒﺷ رﺎﻴﺴﺑ ،ﺪﻳور
.
ﻖﻴﻘﺤﺗ رد ﻲﻓﺮﻃ زا Sagarbieri
ﻦﻳا ﻪﺑ ،نارﺎﻜﻤﻫ و
وﺎﻔﺗ دﻮﺟو ﺎﺑ ﻪﻛ هﺪﺷ هرﺎﺷا ﺐﻠﻄﻣ هزاﺪﻧا ،ﺎﻬﮔﺮﺑ ﻞﻜﺷ رد ت
هﻮﻴﻣ ﻞﻜﺷ و ﮓﻧر ،ﺎﻬﻧآ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ زا ﻪﻧﻮﮔ ﺮﻳز رﺎﻬﭼ يﺎﻫ
(Common, Smyrna, San Pedro, Caprifig) ﻲﺗوﺎﻔﺗ
ﻢﻳﺰﻧآ تﺎﺒﻴﻛﺮﺗ رد ﺖﺳا هﺪﺸﻧ ﻪﻈﺣﻼﻣ ﻚﻴﺘﻟﻮﺌﺗوﺮﭘ يﺎﻫ
) 11 ( ؛
ﻦﻳا ﭗﻴﺗﻮﻨﻓ و ﭗﻴﺗﻮﻧژ ﻦﻴﺑ طﺎﺒﺗرا نداد نﺎﺸﻧ ﺖﻬﺟ لﺎﺣﺮﻫ ﻪﺑ ﻢﻳﺰﻧآ عﻮﻨﺗ ﺮﻈﻧ زا هﺎﻴﮔ ﺎﻫ
ﻪﺑ زﺎﻴﻧ ﺎﻬﻧآ رد دﻮﺟﻮﻣ ﻚﻴﺘﻟﻮﺌﺗوﺮﭘ ي ﻖﻴﻗد تﺎﻌﻟﺎﻄﻣ
ﻲﻣ يﺮﺗ ﺪﺷﺎﺑ .
ﻦﻴﺴﻴﻓ يرﺎﮔﺪﻧﺎﻣ نﺎﻣز رد توﺎﻔﺗ يور ﺮﺑ هﺪﺷ اﺪﺟ يﺎﻫ
ﺎﻬﻧآ ﻲﻟﻮﻜﻟﻮﻣ رﺎﺘﺧﺎﺳ رد توﺎﻔﺗ زا نﺎﺸﻧ ،ﻲﻧﻮﻳ ﺾﻳﻮﻌﺗ نﻮﺘﺳ رد توﺎﻔﺗ رد ﻪﻛ دراد ﻲﻣ ﺲﻜﻌﻨﻣ ﻚﻳﺮﺘﻜﻟاوﺰﻳاpH
دﻮﺷ .
ﭘﺎﭘ ،دﻮﺧ هوﺮﮔﺮﺳ ﺪﻨﻧﺎﻣ ﻢﻳﺰﻧآ ﻦﻳا ﻪﻛ ﺎﺠﻧآ زا ﻦﻳﺎ
) ﻪﺘﺳد زا
ﺎﻫزﺎﺌﺗوﺮﭘ ﻦﻴﺌﺘﻴﺳ (
ﻲﺻﺎﺧ هﺎﮕﻳﺎﺟ ،ﺖﺳا يدﺎﻳز ﺖﻴﻟﺎﻌﻓ ياراد ،
ﻮﻨﻴﻣآ ﻲﻟاﻮﺗ زﻮﻨﻫ نﻮﻨﻛﺎﺗ ﻪﻛ ﻦﻳا و دراد ﻂﺒﺗﺮﻣ ﻊﻳﺎﻨﺻ رد ﻲﻣ ﺖﺳا هﺪﺸﻧ ﺺﺨﺸﻣ ﻞﻣﺎﻛ رﻮﻃ ﻪﺑ نآ يﺎﻫﺪﻴﺳا ﻪﻛ ﺪﺒﻠﻃ
دﺮﻴﮔ ترﻮﺻ ﺎﻬﻧآ يور يﺮﺘﺸﻴﺑ تﺎﻘﻴﻘﺤﺗ هﺪﻨﻳآ رد .
ﻞﻜﺷ 3 - ﺮﻳز رﺎﻬﭼ ﻪﺑاﺮﻴﺷ ماﺮﮔﻮﺗﺎﻣوﺮﻛ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ زا هوﺮﮔ
نﻮﺘﺳ يور ﺮﺑ CM-Cellulose
لوﺪﺟ 1 - ﺮﻴﺠﻧا ﺖﺧرد ﻪﺑاﺮﻴﺷ رد دﻮﺟﻮﻣ يﺎﻫزﺎﺌﺗوﺮﭘ ﺺﻴﻠﺨﺗ ﻪﺻﻼﺧ
ﺐﻴﻛﺮﺗ ﺪﺣاو رد ﺖﻴﻟﺎﻌﻓ ﻢﺠﺣ
ﻢﺠﺣ ) ml±SD ( ﺖﻈﻠﻏ ﻦﻴﺌﺗوﺮﭘ
mg/ml±SD ﻞﻛ ﺖﻴﻟﺎﻌﻓ
ﺖﻴﻟﺎﻌﻓ Unit±SD ﺖﻴﻟﺎﻌﻓ
Unit/mg ﻲﺷﻮﻣﺎﺧ ﺐﻳﺮﺿ
ﻲﻟﻮﻣ ) 4280 (
ﻪﺑاﺮﻴﺷ فﺎﻔﺷ هﺪﺷ 15
±5 242
±3 110 3630 ﺪﺸﻧ ﻦﻴﻴﻌﺗ . ﺪﺸﻧ ﻦﻴﻴﻌﺗ . ﺪﺸﻧ ﻦﻴﻴﻌﺗ .
ءﺰﺟ III
9
±5 41 5 /
±1 7 / 17 370 1 /
±3 6 / 61 11 /
±0 32 / 2 2 /
±0 9 / 30
ءﺰﺟ 15 IV
±4 1 / 36 3 /
±1 3 / 15 543 2 /
±4 7 / 62 16 /
±0 36 / 2 3 /
±0 2 / 21
ءﺰﺟ 20 V
±4 31 5 /
±1 9 / 13 620 1 /
±4 9 / 63 15 /
±0 41 / 2 3 /
±0 5 / 21
ءﺰﺟ 23 VI
±4 37 5 /
±1 6 / 14 851 4 /
±3 2 / 67 13 /
±0 53 / 2 3 /
±0 1 / 22
لوﺪﺟ 2 - ﻪﻧﻮﮔﺮﻳز ﺲﻜﺗﻻ زا هﺪﺷ اﺪﺟ ﻒﻠﺘﺨﻣ ياﺰﺟا ﺖﻴﻟﺎﻌﻓ ﺎﻜﻳرﺎﻛ سﻮﻜﻴﻓ ﻒﻠﺘﺨﻣ يﺎﻫ
(Unit/mg)
ﻪﻧﻮﮔﺮﻳز A
B C D E F G H I J
Black mission
96 / 3 1 / 10 0 0 46 / 2 0 84 / 6 73 / 2 0 96 / 9
Stanford
81 / 2 51 / 5 74 / 5 98 / 5 0 71 / 4 15 / 6 72 / 3 0 85 / 9
0 King
0 21 / 7 6 0 53 / 4 95 / 6 62 / 6 1 / 5 9 / 3
Calimyrna
97 / 2 0 0 0 0 46 / 4 46 / 4 96 / 3 0 0
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680-85.
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7: 163-75.
١١
Extraction and discrimination between four proteases from Khorasan native fig tree
(Ficus carica)
O. Rajabi*, A. Varaste**, J. Danaee Baghaki***, A. Baratian***, L. Jahangiri***
* Assistant Professor, Department of Medicinal Chemistry, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
** Associate Professor, Department of Immunology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
***Student of Pharmacy, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
Abstract
Background and Aim: The protease activity of fig latex has been reported in several articles. This study was carried out aiming at a comparison between proteases of the latex of Khorasan native fig tree and those of carica subspecies.
Materials and Methods: In this experimental study, four latex proteases (Ficins) were extracted using salt fractionation and chromatography on Mono-S column (Ion exchange media). Parameters such as activity, concentration and molar extinction coefficient were determined at least 3 times per sample. The different samples were compared according to activity and molar extinction coefficient, using ANOVA statistical analysis. P<0.05 was taken as significant.
Results: These proteases were similar with regard to molar extinction coefficient, activity and molecular weight. Activity and molecular weight were determined by cosein perpetration and SDS- PAGE respectively. Besides, the enzymes showed different properties as to retention time on cation exchanging column. This is supposed to be because of little differences in their primary structures and peptide maps. Protein concentration and the activity of latex was calculated to be 110 (mg/ml) and 242 (IU/ml) respectively. Agents having enzymatic activity were found in fraction III-VI.
Molecular weight of Ficins III- VI was stimated at about 26.5- 121. A 1% solution of proteins of Fractions III- VI in 0.05 M NH4CO3 had a molar absorption coefficient (ε280 nm) of about 22, 21.
5, 21, 21.9 respectively. Enzymatic activity of Ficins (fractions III-VI) using casein substrate was 61.6, 62.7, 63.4 and 67.2 (unit/mol) respectively.
Conclusion: According to this study, latex of Khorasan native Fig tree is relatively similar to Calimyrna subspecies growing in California/USA among the four subspecies.
Key Words: Fig latex; Ficin; Purification; Enzymatic activity
