شناسايی 5 جهش جديد در ژن گليکوپروتئين Ibα پلاکت در بيماران برنارد- سولير ايران

(1)

ﻲﻳﺎﺳﺎﻨﺷ

5 ﺪﻳﺪﺟ ﺶﻬﺟ

ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد

Ib

α

ﺖﻛﻼﭘ

نارﺎﻤﻴﺑ رد

درﺎﻧﺮﺑ

-ناﺮﻳا ﺮﻴﻟﻮﺳ

يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ I * ﻲﻤﻇﺎﻛﺪﻤﺣاﺮﺘﻛد II يرﻻرﺎﮕﺘﺳرﻢﺳﺎﻗﺮﺘﻛد III ﻼﻋنوﺪﻳﺮﻓﺮﺘﻛد IV نﺎﮔدازلﻮﺳرﻢﻳﺮﻣ V

هﺪﻴﻜﭼ

ﻪﻨﻴﻣز فﺪﻫو : ﺖﺳاﻲﺛراهﺪﻨﻫديﺰﻳﺮﻧﻮﺧيرﺎﻤﻴﺑﺮﻴﻟﻮﺳدرﺎﻧﺮﺑمرﺪﻨﺳ ﻪﻛ ﻟدﻪﺑ ﻲﻟﻮﻜﻟﻮﻣﺺﻳﺎﻘﻧﻞﻴ ﺲﻜﻠﭙﻤﻛ GP Ib-IX-V (GlycoProtein) ﻪﻛﻲﺘﻛﻼﭘ هﺮـﻴﺠﻧزرﺎـﻬﭼزا GP Ibα, GP Ibβ, GP IX, GP V ،ﺖـﺳاهﺪـﺷﻞﻴﻜﺸـﺗ قﺎﻔﺗا ﻲﻣ ﺪﺘﻓا . GP Ibα ﺪﻧﺎﮕﻴﻟﻪﺑلﺎﺼﺗالﻮﺌﺴﻣوهدﻮﺑﺲﻜﻠﭙﻤﻛﻦﻳاهﺮﻴﺠﻧزﻦﻳﺮﺘﮔرﺰﺑ ﻲـﻣ ﺪـﺷﺎﺑ . ﺮﺘﺸـﻴﺑ ﺶـﻬﺟ يﺎـﻫ هﺪﺷﻲﻳﺎﺳﺎﻨﺷ درﺎﻧﺮﺑمرﺪﻨﺳﺎﺑطﺎﺒﺗرارد ﺮﻴﻟﻮـﺳ رد GP Ibα ﺖـﺳاهدﻮـﺑ . وﻲـﺳرﺮﺑﻪـﻌﻟﺎﻄﻣﻦـﻳامﺎـﺠﻧازافﺪـﻫ ﻲﺋﺎﺳﺎﻨﺷ نژﻲﻟﻮﻜﻟﻮﻣﺺﻳﺎﻘﻧ GP Ibα ﻲﻧاﺮﻳاﺮﻴﻟﻮﺳدرﺎﻧﺮﺑنارﺎﻤﻴﺑرد ﻲﻣ ﺪﺷﺎﺑ . شور رﺎﻛ : نارﺎﻤﻴﺑﻲﺳرﺮﺑشورﻪﺑﻪﻌﻟﺎﻄﻣﻦﻳا ) Case series ( وﺪﺷمﺎﺠﻧا يرﺎـﻤﻴﺑﻲﺗﺎﻋﻼﻃاﻚﻧﺎﺑزاهدﺎﻔﺘﺳاﺎﺑ يﺎـﻫ ﻧﺎﻣردهﺪﻨﻫديﺰﻳﺮﻧﻮﺧ ﻲﻠﻴﻓﻮﻤﻫﻊﻣﺎﺟهﺎﮕ ناﺮﻳا 12 رﺎﻤﻴﺑ وﻲﻳﺎﺳﺎﻨﺷ ﺪﻨﺘﻓﺮﮔراﺮﻗﻪﻌﻟﺎﻄﻣدرﻮﻣ . ﻪﻳﺎﭘﺮﺑيرﺎﻤﻴﺑﺺﻴﺨﺸﺗ مﻻﻲﺳرﺮﺑ،ﺖﻛﻼﭘشرﺎﻤﺷﻞﻣﺎﺷﻲﭙﻴﺗﻮﻨﻓتﺎﻌﻟﺎﻄﻣ نﻮﺧ ﻪﺑﻲﻫﺪﺨﺳﺎﭘمﺪﻋوﻲﻄﻴﺤﻣ ﺖﺴـﻴﻧﻮﮔآ ترﻮـﺻﻦﻴﺘﺳﻮﺘﺴـﻳر دﻮﺑﻪﺘﻓﺮﮔ . NA D ونارﺎﻤﻴﺑ ﻲﻧﻮﺧيﺎﻬﻧﻮﺘﺳزا يزﺎﺳاﺪﺟﻪﺘﺴﺑاوداﺮﻓا ﺪﺷ . ﻲﺣاﻮﻧ نژهﺪـﻨﻨﻛﺪﻛ GP Ibα ﻪـﺑ 5 ﻪـﻌﻄﻗ نﺎـﺷﻮﭘﻢﻫ ﺪـﺷمﺎـﺠﻧاﺮـﻴﺜﻜﺗويﺪـﻨﺑﻢﻴﺴـﻘﺗ . شورزا CSGE (Conformation Sensitive Gel Electrophoresis)

هدﺎﻔﺘﺳا ﺪﻳدﺮﮔ و ﺎﺘﻳﺎﻬﻧ ﻲﻟاﻮﺗ لژردﺲﻜﻠﺑودوﺮﺘﻫياراديﺎﻫ CSGE ﺪﻧﺪﺷﻲﻟاﻮﺗﻦﻴﻴﻌﺗ . ﻪﺘﻓﺎﻳ ﺎﻫ : 5 ﺪـﺷﻲﻳﺎﺳﺎﻨﺷﺪﻳﺪﺟﺶﻬﺟ : فﺬـﺣ ACCGGCT ﻳﺎـﺟو ﻲﻨﻳﺰﮕ GGA ﺪـﻴﺗﻮﺌﻠﻛﻮﻧﻦﻴـﺑ يﺎـﻫ 425 -419 ، ﺶﻬﺟ يﺎﻫ ﻪﻄﻘﻧ يا C 709 T ، A 710 G ، T 1759 C فﺬﺣو 20 ﺖﻴﻌﻗﻮﻣردﺪﻴﺗﻮﺌﻠﻛﻮﻧ 1819 -1800 . نﻮﻴﺳﺎﺗﻮﻣ يﺎﻫ ﻲﻳﺎﺳﺎﻨﺷ نﻮﻴـﺳﺎﺗﻮﻣزاماﺪـﻛﺮـﻫياﺮـﺑﺪـﻳدﺮﮔﺖـﺒﺛوﺪـﺷهدﺎﺘـﺳﺮﻓنژﻲﻧﺎـﻬﺟﻚـﻧﺎﺑﻪـﺑهﺪﺷ ﺎـﻫ (RFLP) Restriction

Fragment Length Polymorphism

ﻃ ﺪﺷمﺎﺠﻧاوﻲﺣاﺮ . ﻪﺠﻴﺘﻧ يﺮﻴﮔ : هﺮﻴﺠﻧزنژﻲﻟﻮﻜﻟﻮﻣﺺﻳﺎﻘﻧﻪﻌﻟﺎﻄﻣﻦﻳارد GP Ibα نارﺎـﻤﻴﺑردﺎﻬﺘﻛﻼﭘ BSS راﺮـﻗﻪـﻌﻟﺎﻄﻣدرﻮـﻣ وﺖﻓﺮﮔ 5 ﻪﻛﺪﻳﺪﺟنﻮﻴﺳﺎﺗﻮﻣ ﺪﻳدﺮﮔﻲﻳﺎﺳﺎﻨﺷ،دﻮﺑهﺪﺸﻧﻲﻳﺎﺳﺎﻨﺷتﺎﻌﻟﺎﻄﻣﺮﻳﺎﺳرد . ﺪﻴﻠﻛ ﺎﻫهژاو : 1 -درﺎﻧﺮﺑمرﺪﻨﺳ -ﺮﻴﻟﻮﺳ 2 -ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ Ibα 3 -ﻲﻟاﻮﺗﻦﻴﻴﻌﺗ 4 -ﺶﻬﺟ ﺖﻓﺎﻳردﺦﻳرﺎﺗ : 9 / 3 / 88 شﺮﻳﺬﭘﺦﻳرﺎﺗ ، : 30 / 1 / 89

ﻪﻣﺪﻘﻣ

هﺪﻨﻫديﺰﻳﺮﻧﻮﺧيرﺎﻤﻴﺑﺮﻴﻟﻮﺳـ درﺎﻧﺮﺑمرﺪﻨﺳ ﻲﺛرا ثراﻪﺑبﻮﻠﻐﻣموزﻮﺗاثراﻮﺗﺎﺑًﻻﻮﻤﻌﻣﻪﻛﺖﺳا ﻲﻣ ﺪـﺳر ﺖـﻴﻌﻤﺟردنآعﻮﻴﺷو يﺎـﻫ ردﻚـﻳﻪـﺑﻚـﻳدﺰﻧﻒـﻠﺘﺨﻣ نﻮﻴﻠﻴﻣ ﻲﻣ ﺪﺷﺎﺑ . ) 1 و 2 ( دﻪﺑمرﺪﻨﺳﻦﻳا ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔﻲﻟﻮﻜﻟﻮﻣﺺﻳﺎﻘﻧﻞﻴﻟ GP Ib-IX-V ﻲــﻣدﺎــﺠﻳاﺖــﻛﻼﭘ دﻮــﺷ ﻪــﻛ ﻲــﻠﭘهﺮــﻴﺠﻧزرﺎــﻬﭼزا يﺪـﻴﺘﭙﭘ ,GP Ibβ ,GP Ibα GP IX و GPV هﺪـﺷﻞﻴﻜﺸـﺗ ﻞـﻤﻋﺪـﻧاﺮﺒﻠﻳوناورﻮﺘﻛﺎـﻓياﺮـﺑهﺪﻧﺮﻴﮔناﻮﻨﻋﻪﺑوﺖﺳا ﻲﻣ ﺪﻨﻛ . ) 3 و 4 ( GP Ibα ﻦﻳاهﺮﻴﺠﻧزﻦﻳﺮﺘﮔرﺰﺑ ﺲﻜﻠﭙﻤﻛ وهدﻮﺑ ـﻣﺪـﻧﺎﮕﻴﻟﻪـﺑلﺎﺼـﺗالﻮﺌﺴﻣ ﻲ ﺪـﺷﺎﺑ . ﺶـﻘﻧ GP Ib-IX-V ﺖﻛﻼﭘلﺎﺼﺗا ﺎـﺑهﺪـﻳدﺐﻴـﺳآمﻮﻴﻠﺗوﺪـﻧاﺮـﻳزﻲﺣاﻮﻧﻪﺑﺎﻫ ﺪﻧاﺮﺒﻠﻳوناورﻮﺘﻛﺎﻓﻪﻄﺳاو ﻞﻴﻬﺴﺗو ﺖﻛﻼﭘيزﺎﺳلﺎﻌﻓ ﺎﻫ ﺖﻈﻠﻏرد ﻲﻣﻦﻴﺒﻣوﺮﺗﻦﻴﻳﺎﭘيﺎﻫ ﺪـﺷﺎﺑ . ﺮـﻈﻧﻪـﺑهوﻼـﻋﻪـﺑ ﻲﻣ ﺖـﻛﻼﭘزﺎﺳوﺖﺧﻮﺳوﺪﻴﻟﻮﺗردﺪﺳر ﺪـﺷﺎﺑﻞـﻴﺧدﺎـﻫ نﻮــﭼ ﮔﻦــﻳاردﺺــﻘﻧيارادنارﺎــﻤﻴﺑ رﺎــﭼدﻦﻴﺌﺗوﺮﭘﻮــﻜﻴﻠ هزاﺪﻧاﺶﻳاﺰﻓاوﺖﻛﻼﭘداﺪﻌﺗﺶﻫﺎﻛ ﻲﻣﺖﻛﻼﭘ ﺪﻨﺷﺎﺑ . ) 5 ( ﻦﻳا ﻖﻴﻘﺤﺗ ﺎﺑ هدﺎﻔﺘﺳا زا ﺖﻳﺎﻤﺣ ﻲﻟﺎﻣ هﺎﮕﺸﻧاد ﻲﻜﺷﺰﭘمﻮﻠﻋ و تﺎﻣﺪﺧ ﻲﺘﺷاﺪﻬﺑ ﻲﻧﺎﻣرد ناﺮﻳا رد ﺐﻟﺎﻗ ﺪﻛﻲﺗﺎﻘﻴﻘﺤﺗحﺮﻃ 306 / پ لﺎﺳرد 1386 مﺎﺠﻧا هﺪﻳدﺮﮔ ﺖﺳا . (I ﭘمﻮﻠﻋهﺎﮕﺸﻧاد،يژﻮﻟﻮﺗﺎﻤﻫﺪﺷراسﺎﻨﺷرﺎﻛ ناﺮﻳا،ناﺮﻬﺗ،ناﺮﻳاﻲﻧﺎﻣردﻲﺘﺷاﺪﻬﺑتﺎﻣﺪﺧوﻲﻜﺷﺰ (II ناﺮﻳا،ناﺮﻬﺗ،ناﺮﻳاﻲﻧﺎﻣردﻲﺘﺷاﺪﻬﺑتﺎﻣﺪﺧوﻲﻜﺷﺰﭘمﻮﻠﻋهﺎﮕﺸﻧاد،يژﻮﻟﻮﺗﺎﻤﻫهوﺮﮔرﺎﻴﺸﻧاد *) لوﺆﺴﻣﻒﻟﺆﻣ ( (III ناﺮﻳا،ناﺮﻬﺗ،ناﺮﻳاﻲﻧﺎﻣردﻲﺘﺷاﺪﻬﺑتﺎﻣﺪﺧوﻲﻜﺷﺰﭘمﻮﻠﻋهﺎﮕﺸﻧاد،يژﻮﻟﻮﺗﺎﻤﻫهوﺮﮔرﺎﻳدﺎﺘﺳا (IV ﺼﺨﺗقﻮﻓ ناﺮﻳا،ناﺮﻬﺗ،ناﺮﻳاﻲﻠﻴﻓﻮﻤﻫﻊﻣﺎﺟهﺎﮕﻧﺎﻣرد،ﻲﻨﻴﻟﺎﺑيزﻮﻟﻮﺗﺎﻤﻫﺺ (V ناﺮﻳا،ناﺮﻬﺗ،ناﺮﻳاﻲﻠﻴﻓﻮﻤﻫﻊﻣﺎﺟهﺎﮕﻧﺎﻣرد،ﻲﻤﻴﺷسﺎﻨﺷرﺎﻛ

(2)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻴﻠﮔ نژ رد ﻦﻴﺌﺗوﺮﭘﻮﻜ Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار داﺪـﻌﺗﺶﻫﺎـﻛسﺎﺳاﺮﺑﺮﻴﻟﻮﺳـدرﺎﻧﺮﺑمرﺪﻨﺳﺺﻴﺨﺸﺗ ﺖﻛﻼﭘ هاﺮﻤﻫﺎﻫ ﺎﺑ نﺎـﻣزتﺪـﻣﺶﻳاﺰـﻓا،ﺎـﻫنآهزاﺪـﻧاﺶﻳاﺰﻓا ﺖﻛﻼﭘﻲﻫﺪﺨﺳﺎﭘمﺪﻋويوﺮﻧﻮﺧ ﻘﻣردﺎﻫ وﻦﻴﺘﺳﻮﺘﺴـﻳرﻞـﺑﺎ ﻲﻣترﻮﺻلﺎﻣﺮﻧيﺎﻤﺳﻼﭘندوﺰﻓاﺎﺑنآﺢﻴﺤﺼﺗمﺪﻋ ،دﺮـﻴﮔ تاﺪﻫﺎﺸــﻣﺮﻴﻟﻮــﺳـ درﺎــﻧﺮﺑمرﺪﻨــﺳﺺﻴﺨﺸــﺗﺪــﻴﻳﺄﺗرد ﺖﺴﺗمﺎﺠﻧاﺎﺑﻲﺘﺴﻳﺎﺑيﺮﺘﻣﻮﮕﻳﺮﮔا ﻞﻣﺎـﺷﻲﻳﺎﻴﻤﻴـﺷﻮﻴﺑيﺎـﻫ ناﺰﻴﻣيﺮﻴﮔهزاﺪﻧا GP Ib-IX-V هﺮﻴﺠﻧزو هﺪﻨﻫدﻞﻴﻜﺸﺗيﺎﻫ ﻳاﺎﻳويﺮﺘﻣﻮﺘﻴﺳﻮﻠﻓﻂﺳﻮﺗﺖﻛﻼﭘﺢﻄﺳردنآ ردوتﻼﺑﻮﻨﻤ ﺪـﻴﻳﺄﺗﻲﻜﻴﺘﻧژتﺎﻌﻟﺎﻄﻣﺎﺑﺖﻳﺎﻬﻧ لﻼﺘـﺧاﻞـﻣﺎﻋًﺎـﻘﻴﻗدﺎﺗدﻮـﺷ دﻮﺷﻦﻴﻴﻌﺗﺖﻛﻼﭘيدﺮﻜﻠﻤﻋ . ) 5 و 6 ( نارﺎـﻤﻴﺑيوﺮـﺑﻲﻜﻴﺘﻧژ ﻪﻌﻟﺎﻄﻣﭻﻴﻫ نﻮﻨﻛﺎﺗناﺮﻳارد دﻮـﺑهﺪﺸـﻧمﺎـﺠﻧاﺮﻴﻟﻮـﺳـدرﺎﻧﺮﺑ . ﻦـﻳاﻲـﻜﻴﺘﻧژﻪـﻌﻟﺎﻄﻣ ،ﻦﻴﻠﻗﺎــﻧﻦﻴـﻴﻌﺗ،يرﺎـﻤﻴﺑﻲــﻌﻄﻗﺺﻴﺨﺸـﺗﺖـﻬﺟنارﺎـﻤﻴﺑ ﻮﺗزاﻞﺒﻗﺺﻴﺨﺸﺗ ونژﻲﻧﺎـﻬﺟﻚـﻧﺎﺑﻞـﻴﻤﻜﺗﻪﺑﻚﻤﻛ،ﺪﻟ يروﺮـﺿهﺪـﻨﻳآردﺺﻗاﻮﻧﻦﻳاﻪﻌﻟﺎﻄﻣياﺮﺑﻲﺷورﻪﺋارا ﺮـﻈﻧﻪﺑ ﻲـﻣ ﺪﻴـﺳر . هﺮـﻴﺠﻧزﻪـﻛﺎـﺟنآزا GP Ibα ﺶـﻘﻧ رادهﺪــﻬﻋارﺪــﻧﺎﮕﻴﻟﻪــﺑلﺎﺼــﺗا ﻲــﻣ ﻦﻳﺮﺘﺸــﻴﺑوﺪــﺷﺎﺑ نﻮﻴﺳﺎﺗﻮﻣ يﺎﻫ ﺖﺳاهدﻮﺑهﺮﻴﺠﻧزﻦﻳاردهﺪﺷشراﺰﮔ ﻪﺑ مﺎﮔﻦﻴﻟواناﻮﻨﻋ ، ﻮـﻜﻟﻮﻣﺺﻗاﻮﻧﻲﺳرﺮﺑ هﺮـﻴﺠﻧزﻲﻟ GP Ibα نارﺎﻤﻴﺑﻦﻳارد ار ﻢﻳدﺮﻛعوﺮﺷ .

رﺎﻛ

شور

يرﺎﻤﻴﺑﻲﺗﺎﻋﻼﻃاﻚﻧﺎﺑزاﻦﺘﻓﺮﮔﻚﻤﻛﺎﺑ يﺎـﻫ يﺰﻳﺮﻧﻮـﺧ مرﺪﻨــﺳيارادنارﺎــﻤﻴﺑ ،ﻲﻠﻴﻓﻮــﻤﻫﻊﻣﺎــﺟهﺎــﮕﻧﺎﻣردهﺪــﻨﻫد درﺎﻧﺮﺑ داﺪﻌﺗوﻲﻳﺎﺳﺎﻨﺷﺮﻴﻟﻮﺳ 12 ﺪـﻳدﺮﮔبﺎـﺨﺘﻧارﺎﻤﻴﺑ . شرﺎﻤـﺷﻞﻣﺎـﺷﻲﭙﻴﺗﻮﻨﻓتﺎﻌﻟﺎﻄﻣﻪﻳﺎﭘﺮﺑيرﺎﻤﻴﺑﺺﻴﺨﺸﺗ ﭘ ﻣﻲﺳرﺮﺑ،ﺖﻛﻼ ﻮ ﻲﻄﻴﺤﻣمﻻﻚﻳژﻮﻟﻮﻓر و ﻲﻫﺪﺨـﺳﺎﭘمﺪﻋ ﺖــﻛﻼﭘ ﺎــﻫ لﺎــﻣﺮﻧيﺎﻤــﺳﻼﭘرﻮﻀــﺣردﻦﻴﺘﺳﻮﺘﺴــﻳرﻪــﺑ دﻮﺑﻪﺘﻓﺮﮔترﻮﺻ . ناﺰﻴﻣ،ﺎﻬﻧآهداﻮﻧﺎﺧونارﺎﻤﻴﺑزا 15 ﻲﻠﻴﻣ دﺎـﻘﻌﻧاﺪـﺿردنﻮﺧﺮﺘﻴﻟ Na2EDTA (Ethylene Diamine

Tetra Acetic Acid)

وﺪـﺷيروآﻊـﻤﺟ DNA ﺎـﺑﻲﻣﻮـﻧژ شورزاهدﺎﻔﺘــﺳا زﺎــﻨﻴﺌﺗوﺮﭘﻲﺘــﺳد K ــﺳ، ﺪ ﻞﻴــﺳدودﻢﻳ لﻮﺒﻠﮔﺰﻴﻟﺮﻓﺎﺑوعﺎﺒﺷامﻮﻴﻧﻮﻣآتﺎﺘﺳا،تﺎﻔﻟﻮﺳ يﺎﻫ ﺪﻴﻔـﺳ لﻮﺒﻠﮔزا يﺎﻫ ﺪﻳدﺮﮔجاﺮﺨﺘﺳاﺪﻴﻔﺳ . مﺎـﻤﺗﻪـﻜﻨﻳاﻪـﺑﻪﺟﻮﺗﺎﺑ هﺮﻴﺠﻧزهﺪﻨﻨﻛﺪﻛﻲﻟاﻮﺗ GP Ibα راﺮـﻗنوﺰـﮔاﻚﻳنوردرد ﻪﺑدرادﻲﮔرﺰﺑهزاﺪﻧانوﺰﮔاﻚﻳﻦﻳاوﻪﺘﺷاد 5 ﻢﻴﺴﻘﺗﻪﻌﻄﻗ درﻮــﻣوﺪــﺷ ﺖــﻓﺮﮔراﺮــﻗﻪــﻌﻟﺎﻄﻣ . مﺎــﺠﻧاياﺮــﺑ PCR

(Polymerase Chain Reaction)

شورزا Touch down ﺪﺷهدﺎﻔﺘﺳا . ﺮﻤﻳاﺮﭘ يﺎﻫ ﻦﻳاياﺮﺑهدﺎﻔﺘﺳادرﻮﻣ 5 ردﻪـﻌﻄﻗ لوﺪﺟ هرﺎﻤﺷ 1 ﺖﺳاهﺪﺷهدروآ . هرﺎﻤﺷلوﺪﺟ 1 -هﺪﺷهدﺎﻔﺘﺳايﺎﻫﺮﻤﻳاﺮﭘﻲﻟاﻮﺗ PRIMER PRIMER POSITION PRIMER SEQUENCE(5'-3') gagagaaggacgg agtcgag 2780 -2761 1 A ggttgtgtctttcggc aggt 3199 -3218 R 1 A ctgtgaggtctccaa agtgg 3144 -3125 2 A tagccagactgagct tctcc 3566 -3585 R 2 A aaggcaatgagctg aagacc 3530 -3511 3 A cttgtgttggatgca aggag 4087 -4108 R 3 A tccactgcttctctag acag 4075 -4056 4 A ggctgatcaagttca gggat 4473 -4492 R 4 A cacaagcctgatcac tccaa 4413 -4394 5 A ttctctcaaggtcccc aaac 4959 -4978 R 5 A CSGE : سﺎﺳا CSGE ﺲﻜﻠﺑودوﺮﺘﻫﻞﻴﻜﺸﺗيﺎﻨﺒﻣﺮﺑ DNA يارادهﺮــﻴﺠﻧزﻲﻓدﺎﺼــﺗنﺪــﺷﺖــﻔﺟزاﻲــﺷﺎﻧ ﺖـﺳاﻢﻟﺎــﺳهﺮـﻴﺠﻧزﺎــﺑنﻮﻴـﺳﺎﺗﻮﻣ . ياﺮــﺑشورﻦــﻳازا ﺷ ﻨ نﻮﻴﺳﺎﺗﻮﻣ ﻪﻴﻟواﻲﻳﺎﺳﺎ ﺎﻫ دﺎﻔﺘﺳا ﻪـﻠﺣﺮﻣودﻪـﻛﺪـﺷه تﺪـﻣﻪـﺑﺐـﻴﺗﺮﺗ ﻪـﺑلﺎﺼﺗاوﺐﻴﻠﻘﺗ 5 ﻪـﻘﻴﻗد C ° 98 و 30 ﻪﻘﻴﻗد C ° 65 ﻲـﻣ ﺪـﺷﺎﺑ . لژرد CSGE ﻦﻠﻴـﺗا،ﺪـﻴﻣﺎﻣﺮﻓزا ولﻮﻜﻴﻠﮔ BAP (1, 4-BisAcrolyl piperazine) ﺑ ﻪ رﻮﻈﻨﻣ ﺪﺷهدﺎﻔﺘﺳاﺖﻴﺳﺎﺴﺣﺶﻳاﺰﻓا . لژردﻪﻛﻲﺗﺎﻌﻄﻗ CSGE ﺪﺷﻲﻟاﻮﺗﻦﻴﻴﻌﺗ،ﺪﻧدﻮﺑﺲﻜﻠﺑودوﺮﺘﻫياراد ﺪﻧ اﻪـﻛ ﺪـﺘﻣز رﻮـﻈﻨﻣﻦـﻳاﻪـﺑﺪـﺷهدﺎﻔﺘـﺳاﺮﺠﻧﺎـﺳ 2 رﺎـﺑ PCR مﺎـﺠﻧا ،ﺪﺷ PCR ﻚﻳلوا PCR ﺺﻟﺎـﺧلﻮﺼﺤﻣﻪﻛدﻮﺑﻲﻟﻮﻤﻌﻣ ياﺮﺑنآهﺪﺷيزﺎﺳ PCR ﺲﻧﺎﻜـﺳصﻮﺼـﺨﻣﻪـﻛمود

(3)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار ﺖــﻓﺰﮔراﺮــﻗهدﺎﻔﺘــﺳادرﻮــﻣﺖــﺳا . مﺎــﺠﻧاياﺮــﺑ PCR و وﺎﻤﮕﻴــﺳﺖﻛﺮــﺷﺖــﻴﻛزانآلﻮﺼــﺤﻣيزﺎــﺳﺺﻟﺎــﺧ ﺪــﺷهدﺎﻔﺘــﺳانژﺎــﻴﻛ . زاﻲﻟاﻮــﺗﻦﻴــﻴﻌﺗياﺮــﺑ هﺎﮕﺘــﺳد Pharmasia Amersham ﻲــــﺳرﺮﺑياﺮــــﺑو ﻲﻟاﻮــــﺗ ﺪــﻴﺗﻮﺌﻠﻛﻮﻧ ﺎــﻫ زا مﺮــﻧ راﺰــﻓا

ALF express sequence

analysis ﺪﻳدﺮﮔهدﺎﻔﺘﺳا . RFLP : زاهدﺎﻔﺘﺳاﺎﺑ مﺮـﻧ راﺰـﻓا Gene Runner يﻮـﮕﻟا، ﻢﻳﺰﻧآﻂﺳﻮﺗشﺮﺑتاﺮﻴﻴﻐﺗ يﺎﻫ ﻢﻳﺰﻧآ،ﻲﺳرﺮﺑهﺪﻨﻨﻛدوﺪﺤﻣ ﻚﻴﻨﻜﺗوبﺎﺨﺘﻧاﺐﺳﺎﻨﻣ RFLP ياﺮﺑ نﻮﻴـﺳﺎﺗﻮﻣ ـﻫ يﺎ ﻪـﺘﻓﺎﻳ ﺪﺷمﺎﺠﻧاوﻲﺣاﺮﻃهﺪﺷ . نﻮﻴـﺳﺎﺗﻮﻣرﻮﻀﺣ ﻲـﻣ ﺎـﺑﺪـﻧاﻮﺗ ﺪﻫدﺮﻴﻴﻐﺗارشﺮﺑيﻮﮕﻟا،نژﻲﻟاﻮﺗﺮﻴﻴﻐﺗ شورﻦـﻳازاﻪـﻛ ﺪﺷهدﺎﻔﺘﺳانﻮﻴﺳﺎﺗﻮﻣرﻮﻀﺣﺪﻴﻳﺎﺗوﻲﺑﺎﻳدرياﺮﺑ .

ﻪﺘﻓﺎﻳ

ﺎﻫ

ﻪﻌﻄﻗردرﺎﻤﻴﺑﻚﻳ 2 ﻪﻌﻄﻗردرﺎﻤﻴﺑود، 3 رﺎـﻤﻴﺑودو ﻪﻌﻄﻗرد ﻢﻫ 5 لژرد ﺲﻜﻠﺑودوﺮﺘﻫياراد CSGE ﺪـﻧدﻮﺑ ﺪﻧﺪـﺷ ﻲﻟاﻮﺗﻦﻴﻴﻌﺗ ﻪﻛ . ﻲﻟاﻮـﺗﻦـﻳاﻲـﺳرﺮﺑ ﺎـﻫ ﻲﻟاﻮـﺗﺎـﺑ رﻮﻀﺣ،لﺎﻣﺮﻧ 5 ﺶـﻬﺟ ﺪـﻳﺪﺟ نژرد GP Ibα ار نﺎﺸـﻧ ﻪﻛداد ﻦﻳا ﺶﻬﺟ ﺎﻫ ) هرﺎﻤـﺷﻞﻜﺷ 1 ( ﻞﻣﺎـﺷ ﺮـﻳزدراﻮـﻣ ﻲــﻣ ﺪــﺷﺎﺑ : فﺬــﺣ ACCGGCT ﻲﻨﻳﺰﮕﻳﺎــﺟو GGA ﻦﻴــﺑ ﺪﻴﺗﻮﺌﻠﻛﻮﻧ يﺎﻫ 425 -419 ، ﺶﻬﺟ يﺎـﻫ ﻪـﻄﻘﻧ يا C 709 T ، A 710 G ، T 1759 C فﺬﺣو 20 ﺖﻴﻌﻗﻮﻣردﺪﻴﺗﻮﺌﻠﻛﻮﻧ 1819 -1800 . ﺶﻬﺟ يﺎﻫ هﺪﺷﻲﻳﺎﺳﺎﻨﺷ شورﻪﻠﻴـﺳﻮﺑ RFLP و ﻢﻳﺰـﻧآزاهدﺎﻔﺘـﺳاﺎـﺑﺐﻴﺗﺮﺗﻪﺑ يﺎـﻫ هﺪـﻨﻨﻛدوﺪﺤﻣ BsiYI ، MspI ، ECORII ، TaqI و BsiYI ﺪﻧﺪﺷﺪﻴﻳﺎﺗوﻲﺳرﺮﺑ .

(4)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار هرﺎﻤﺷﻞﻜﺷ 1 -رﻮﻀﺣ 5 ﺶﻬﺟ ﺪﻳﺪﺟ نژرد GP Ibα نﻮﻴﺳﺎﺗﻮﻣ ACCGGCT/Ins GGA Del ﺑ فﺬـﺣﺚﻋﺎ شﺮــﺑﻞــﺤﻣﻚــﻳ ﻢﻳﺰــﻧآ BsiYI ﺖــﻴﻌﻗﻮﻣرد 321 ﻪــﻌﻄﻗ 2 ﻲﻣ دﻮﺷ . ،ﻢﻟﺎﺳﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢﻀﻫ 4 يﺎﻫهزاﺪﻧاﻪﺑﻪﻌﻄﻗ bp 48 ، bp 45 ، bp 140 ، bp 227 ﻪــﻌﻄﻗﻲــﻤﻳﺰﻧآﻢﻀــﻫﻪــﺠﻴﺘﻧو ﺖـــــﻧﺎﺗﻮﻣ 3 ﻪـــــﻌﻄﻗ bp 48 ، bp 185 و bp 227 ﻲـــــﻣ ﺪـــــﺷﺎﺑ ) ﻞﻜﺷ هرﺎﻤﺷ 2 .( ﺎﺑ زرﻮـﻓوﺮﺘﻜﻟاﺖـﻛﺮﺣردﻪﻜﻨﻳاﻪﺑﻪﺟﻮﺗ ﻪﻌﻄﻗ bp 45 و bp 48 ﺖﻛﺮﺣﻢﻫﺎﺑ ﻲﻣ ﺪﻨﻨﻛ ، رﺎـﻈﺘﻧا ﻲـﻣ رددور ﻢﻟﺎــﺳﻪــﻌﻄﻗزرﻮــﻓوﺮﺘﻜﻟالژ 3 ﺪــﻧﺎﺑ bp 48 ، bp 140 ، bp 227 ودﻮﺷهﺪﻫﺎﺸﻣ ﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗرد 3 ﺪﻧﺎﺑ bp 48 ، bp 185 و bp 227 . هﺪﺸﻧﻢﻀﻫﻪﻌﻄﻗلﻮﻃ 461 ﻲﻣزﺎﺑﺖﻔﺟ ﺪﺷﺎﺑ . ﻞﻜﺷ هرﺎﻤﺷ 2 نﻮﻴﺳﺎﺗﻮﻣيارادﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢﻀﻫ ACCGGCT

del / GGA ins

ﻂﺳﻮﺗ . BsiYI هرﺎﻤﺷ 1 هرﺎﻤـﺷ،هﺪﺸﻧﻢﻀﻫلﻮﺼﺤﻣ 2 هرﺎﻤﺷ،ﻢﻟﺎﺳ 3 رﺎﻤﻴﺑ ) تﻮﮕﻳزﻮﻤﻫ ( هرﺎﻤﺷ، 4 رﺎﻤﻴﺑﺪﻟاو ) تﻮﮕﻳزوﺮﺘﻫ ( هرﺎﻤﺷو 5 ﺮﻛرﺎﻣ ﻲﻣ ﺪﺷﺎﺑ . ﻪﻌﻄﻗ bp 48 رﻮﻃنﺎﻤﻫوﻪﺘﻓرنوﺮﻴﺑلژزا هﺪﻫﺎﺸﻣﻪﻛ ﻲﻣ ﻪﻌﻄﻗوديارادرﺎﻤﻴﺑدﺮﻓدﻮﺷ 227 و 185 لﺎﻣﺮﻧدﺮﻓو ﻪﻌﻄﻗودياراد 227 و 140 يزﺎﺑﺖﻔﺟ ﻲﻣ ﺪﺷﺎﺑ . نﻮﻴﺳﺎﺗﻮﻣ C 709 T شﺮﺑﻞﺤﻣﻚﻳدﺎﺠﻳاﺚﻋﺎﺑ ﻢﻳﺰﻧآياﺮﺑ MspI ﺖﻴﻌﻗﻮﻣرد 224 ﻪﻌﻄﻗرد 3 ﻲﻣ دﻮﺷ . ﻲﻟﺎﺣرد ﻪـﻌﻄﻗﻪﻛ دراﺪـﻧدﻮـﺟوﻢﻳﺰﻧآﻦﻳاياﺮﺑﻲﺷﺮﺑﻞﺤﻣﻢﻟﺎﺳ . ﺎـﺑﻦﻳاﺮﺑﺎـﻨﺑ ﻪﻌﻄﻗﻚﻳﻢﻟﺎﺳﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢﻀﻫ 597 ﻞﺻﺎﺣ ﻲﻣ دﻮـﺷ . رد ﻪـﻌﻄﻗودﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢﻀﻫﺎﺑﻪﻛﻲﻟﺎﺣ 224 و 373 ﻞﺻﺎﺣ ﻲﻣ دﻮﺷ ) هرﺎﻤﺷﻞﻜﺷ 3 .( ﻞﻜﺷ هرﺎﻤﺷ 3 نﻮﻴـﺳﺎﺗﻮﻣ يارادﻪـﻌﻄﻗﻲـﻤﻳﺰﻧآﻢﻀﻫ C 709 T ﻂﺳﻮﺗ 1 Msp . هرﺎﻤﺷ 1 هرﺎﻤﺷ،هﺪﺸﻧﻢﻀﻫلﻮﺼﺤﻣ 2 دﺮـﻓﻪﻧﻮﻤﻧ هرﺎﻤﺷ،ﻢﻟﺎﺳ 3 رﺎﻤﻴﺑدﺮﻓﻪﻧﻮﻤﻧ ) تﻮﮕﻳزﻮﻤﻫ ( نﺎﺸﻧار ﻲﻣ ﺪﻫد . لﻮﻃ ﻪﻌﻄﻗ 597 ددﺮـﻓردنآﻲـﻤﻳﺰﻧآﻢﻀـﻫ زاﻪـﻛﺖﺳازﺎﺑﺖﻔﺟ يارا ﻪﻌﻄﻗودنﻮﻴﺳﺎﺗﻮﻣ 373 و 224 ﻞﺻﺎﺣ ﻲـﻣ ردﻪـﻛﻲﻟﺎـﺣرددﻮـﺷ ﻪﻌﻄﻗﻢﻟﺎﺳدﺮﻓ 597 ﻲﻗﺎﺑهدرﻮﺨﻧﺖﺳديزﺎﺑﺖﻔﺟ ﻲﻣ ﺪﻧﺎﻣ . نﻮﻴـﺳﺎﺗﻮﻣ A 710 G شﺮـﺑﻞـﺤﻣﻚـﻳفﺬـﺣﺚـﻋﺎﺑ EcorП ﻪﻌﻄﻗرد 3 ) 222 ( ﻲﻣ دﻮﺷ . ﻪـﻌﻄﻗ ﻲﻤﻳﺰﻧآ ﻢﻀﻫزا ﻢﻟﺎــﺳ 5 ﻪــﻌﻄﻗ 26 ، 75 ، 139 ، 161 و 196 يزﺎــﺑﺖــﻔﺟ

(5)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار ﻞﺻﺎﺣ ﻲﻣ ﻀﻫزاودﻮﺷ ﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢ 4 ﻪﻌﻄﻗ 26 ، 75 ، 139 و 357 ) هرﺎﻤﺷﻞﻜﺷ 4 .( ﻪﻌﻄﻗ يﺎﻫ 26 و 75 دﺮﻓرد،ﻪﺘﻓرنوﺮﻴﺑلژزازرﻮﻓوﺮﺘﻜﻟامﺎﮕﻨﻫيزﺎﺑﺖﻔﺟ تﺎﻌﻄﻗﻢﻟﺎﺳ 196،161 و 139 هﺪﻫﺎﺸﻣ ﻲﻣ ﻲﻟﺎـﺣرددﻮﺷ تﺎﻌﻄﻗرﺎﻤﻴﺑدﺮﻓردﻪﻛ 357 و 139 هﺪﻫﺎﺸﻣيزﺎﺑﺖﻔﺟ ﻲﻣ دﻮﺷ . ﻞﻜﺷ هرﺎﻤﺷ 4 ﻲـﻤﻳﺰﻧآﻢﻀﻫ نﻮﻴـﺳﺎﺗﻮﻣيارادﻪـﻌﻄﻗ A 710 G ﻂﺳﻮﺗ EcorП . هرﺎﻤﺷ 1 هرﺎﻤـﺷ،هﺪﺸـﻧﻢﻀﻫﻪﻧﻮﻤﻧ 2 دﺮـﻓﻪـﻧﻮﻤﻧ هرﺎﻤـــﺷ،ﻢﻟﺎـــﺳ 3 و 4 رﺎـــﻤﻴﺑﻪـــﻧﻮﻤﻧ ﻲـــﻣ هرﺎﻤـــﺷوﺪﻨـــﺷﺎﺑ 5 ﺮﻛرﺎﻣ . ﺪﻧﺎﺑ يﺎﻫ 26 و 75 ردﻪﺠﻴﺘﻧردوﻪﺘﻓرنوﺮﻴﺑلژزايزﺎﺑﺖﻔﺟ يﺎﻫﺪﻧﺎﺑﻢﻟﺎﺳﻪﻌﻄﻗ 196 ، 161 و 139 ﺪﻧﺎﺑﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗويزﺎﺑﺖﻔﺟ يﺎﻫ 357 و 139 هﺪﻫﺎﺸﻣيزﺎﺑ ﻲﻣ دﻮﺷ . نﻮﻴﺳﺎﺗﻮﻣ T 1759 C شﺮـﺑﻞـﺤﻣﻚﻳفﺬﺣﺚﻋﺎﺑ 1 Taq رد ﻪﻌﻄﻗ 5 ﻲﻣ دﻮﺷ . 1 Taq ردارﻢﻟﺎﺳﻪﻌﻄﻗ 391 ﻲﻟوهدادشﺮﺑ ﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗردشﺮﺑﻪﺑردﺎﻗ ﻲﻤﻧ ﺪﺷﺎﺑ ) هرﺎﻤﺷﻞﻜﺷ 5 .( ﻞﻜــﺷ هرﺎﻤــﺷ 5 نﻮﻴــﺳﺎﺗﻮﻣيارادﻪــﻌﻄﻗﻲــﻤﻳﺰﻧآﻢﻀــﻫ T 1759 C ﻂﺳﻮﺗ 1 Taq نﺎﺸﻧار ﻲﻣ ﺪﻫد . هرﺎﻤﺷ 1 هرﺎﻤﺷ،هﺪﺸﻧﻢﻀﻫﻪﻧﻮﻤﻧ 2 ﻪﻧﻮﻤﻧ هرﺎﻤﺷ،ﻢﻟﺎﺳدﺮﻓ 3 رﺎﻤﻴﺑدﺮﻓﻪﻧﻮﻤﻧ ) تﻮﮕﻳزﻮﻤﻫ ( هرﺎﻤﺷ، يﺎـﻫ 4 و 5 ﻪـﻧﻮﻤﻧ رﺎﻤﻴﺑﻦﻳﺪﻟاو ﻲﻣ ﺪﻨﺷﺎﺑ . ﺪـﻧﺎﺑوديارادﻢﻟﺎـﺳدﺮﻓ 391 و 194 يزﺎـﺑﺖـﻔﺟ ﺪﻧﺎﺑيارادﺎﻬﻨﺗرﺎﻤﻴﺑﻪﻛﻲﻟﺎﺣردهدﻮﺑ 585 ﻲـﻣ ﺪـﺷﺎﺑ . تﻮـﮕﻳزوﺮﺘﻫداﺮـﻓا ) ﻦﻳﺪﻟاو ( ﺪﻧﺎﺑﻪﺳﺮﻫ 391،585 و 194 ﺪﻨﺘﺴﻫاراداريزﺎﺑﺖﻔﺟ . فﺬــﺣﺎــﺑ 20 ﺪــﻴﺗﻮﺌﻠﻛﻮﻧ ﺖــﻴﻌﻗﻮﻣرد 1819 -1800 ، ﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗلﻮﻃ 565 ﺪـﻴﺗﻮﺌﻠﻛﻮﻧ ﻲـﻣ ﺚـﻋﺎﺑﻪـﻛدﻮـﺷ ﻢﻳﺰﻧآيﻮﮕﻟاﺮﻴﻴﻐﺗ 1 Bsiy ﻲﻣ دﻮﺷ . ﻞـﺤﻣودﻢﻟﺎـﺳﻪﻌﻄﻗ ﺖــﻴﻌﻗﻮﻣردشﺮــﺑ يﺎــﻫ 187 و 375 ﻢﻀــﻫزاﻪــﻛدراد نآﻲﻤﻳﺰﻧآ 3 ﻪﻌﻄﻗ 187 ، 188 و 210 ﻞـﺻﺎﺣ ﻲـﻣ ـﺷ دﻮ . ﺖﻴﻌﻗﻮﻣردشﺮﺑﻞﺤﻣﻪﺳﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗ يﺎـﻫ 433 ، 375 و 187 ﻪﻌﻄﻗرﺎﻬﭼنآﻲﻤﻳﺰﻧآﻢﻀﻫزاﻪﻛدراد 58 ، 187 ، 188 و 132 ﻞﺻﺎﺣ ﻲﻣ دﻮﺷ ) هرﺎﻤـﺷﻞﻜﺷ 6 .( ﻪـﻌﻄﻗ 187 و 188 رﺎـﻈﺘﻧاوهدﻮﺒﻧﻢﻫزاﻚﻴﻜﻔﺗﻞﺑﺎﻗ ﻲـﻣ ﻪـﻌﻄﻗدور ﺪﻧﺎﺑ ودﻢﻟﺎﺳ 187 و 210 ﺖﻧﺎﺗﻮﻣﻪﻌﻄﻗوﺪﺷﺎﺑﻪﺘﺷاد 3 ﺪﻧﺎﺑ 58 ، 132 و 187 ﺪﺷﺎﺑﻪﺘﺷاد . زرﻮـﻓوﺮﺘﻜﻟامﺎﮕﻨﻫﻪﺑ ، ﻪﻌﻄﻗ 58 ودرﺎـﻤﻴﺑرد،ﻪـﺘﻓرنوﺮـﻴﺑلژزايزﺎـﺑﺖـﻔﺟ ﻪﻌﻄﻗ 132 و 187 ﻪـﻌﻄﻗودلﺎﻣﺮﻧردويزﺎﺑﺖﻔﺟ 187 و 210 هﺪﻫﺎﺸﻣيزﺎﺑﺖﻔﺟ ﻲﻣ دﻮﺷ . ﻞﻜﺷ هرﺎﻤﺷ 6 يارادﻪﻌﻄﻗﻲﻤﻳﺰﻧآﻢﻀﻫ ﺣ فﺬ 20 ارﺪﻴﺗﻮﺌﻠﻛﻮﻧ ﻂﺳﻮﺗ 1 Bsiy نﺎﺸﻧ ﻲﻣ ﺪﻫد . هرﺎﻤﺷ 1 ﻮﻤﻧ هرﺎﻤـﺷ،هﺪﺸـﻧﻢﻀﻫﻪﻧ 2 هرﺎﻤﺷ،لﺎﻣﺮﻧ 3 هرﺎﻤﺷورﺎﻤﻴﺑ 4 ﺮﻛرﺎـﻣ ﻲـﻣ ﺪـﺷﺎﺑ . ودلﺎـﻣﺮﻧرد ﻪﻌﻄﻗ 188 و 210 ﻪﻌﻄﻗودرﺎﻤﻴﺑردويزﺎﺑﺖﻔﺟ 132 و 188 ﺖﻔﺟ هﺪﻫﺎﺸﻣيزﺎﺑ ﻲﻣ دﻮﺷ .

ﺚﺤﺑ

ﻪﺠﻴﺘﻧ

و

يﺮﻴﮔ

5 ﺪـﻳﺪﺟ ﻪـﻌﻟﺎﻄﻣﻦـﻳاردهﺪـﺷﻲﻳﺎﺳﺎﻨﺷ نﻮﻴﺳﺎﺗﻮﻣ شراﺰـــﮔرﺎـــﺑﻦﻴـــﻟواياﺮـــﺑوهدﻮـــﺑ ﻲـــﻣ دﻮـــﺷ . فﺬـــــﺣ ACCGGCT ﻲﻨﻳﺰﮕﻳﺎـــــﺟو GGA ﻦﻴـــــﺑ ﺪﻴﺗﻮﺌﻠﻛﻮﻧ يﺎﻫ 425 -419 جرﺎﺧ ﻦﻣودهﺪﻨﻨﻛﺪﻛﻪﻴﺣﺎﻧ رد ﺖﺳاهدﺎﺘﻓاقﺎﻔﺗاﻲﻟﻮﻠﺳ . فﺬـﺣﻪﺑﻪﺟﻮﺗﺎﺑ 7 ﺪـﻴﺗﻮﺌﻠﻛﻮﻧ

(6)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار ﻲﻨﻳﺰﮕﻳﺎﺟو 3 ﻲﻟاﻮﺗﺐﻟﺎﻗﺮﮕﻳدﺪﻴﺗﻮﺌﻠﻛﻮﻧ يﺎﻫ هﺪـﻨﻨﻛﺪـﻛ هﺮﻴﺠﻧزياﺮﺑ GP Ibα زاﺲـﭘهﺮـﻴﺠﻧزوهدﺮـﻛﺮﻴﻴﻐﺗ 21 ﺎﻬﺘﻧاﻪﺑﻪﻨﻴﻣآﺪﻴﺳا ﻲﻣ ﺪﺳر . ﺖـﻧﺎﺗﻮﻣهﺮﻴﺠﻧز 130 ﺪﻴـﺳا ﻢﻟﺎـﺳهﺮﻴﺠﻧزﻪﺑﺖﺒﺴﻧوﻪﺘﺷادﻪﻨﻴﻣآ 480 ﻪـﻨﻴﻣآﺪﻴـﺳا درادلﻮﻃﺮﺘﻤﻛ . ،هدﻮـﺑﺺﻗﺎﻧهﺮﻴﺠﻧزﻦﻳاﻪﺠﻴﺘﻧرد ﻪـﺑو ﺮﻈﻧ ﻲﻣ ﺪﺳر ﺎـﺑطﺎـﺒﺗرايراﺮـﻗﺮﺑﻪـﺑردﺎﻗ،ﺰﺘﻨﺳزاﺲﭘ هﺮﻴﺠﻧزﺮﻳﺎﺳ ﺎﻫ ﺲﻜﻠﭙـﻤﻛيﺮﻴﮔﻞﻜﺷمﺪﻋﺚﻋﺎﺑوهدﻮﺒﻧ لﻮﻠﺳﺢﻄﺳرد ﻲﻣ دﻮﺷ . ﺚﻋﺎﺑنﻮﻴﺳﺎﺗﻮﻣﻦﻳا ﻚﻳ فﺬﺣ ﻢﻳﺰﻧآياﺮﺑشﺮﺑﻞﺤﻣ Bsiy1 ﻲﻣ ﻪﻛددﺮﮔ ﻲﻣ ﻪـﻳﺎﭘ ﺪﻧاﻮﺗ ﻲﺣاﺮﻃ RFLP ﺪﺷﺎﺑنﻮﻴﺳﺎﺗﻮﻣﻦﻳاﻲﺑﺎﻳدرياﺮﺑ . نﻮﻴﺳﺎﺗﻮﻣ C 709 T نوﺪﻛﺮﻴﻴﻐﺗﺚﻋﺎﺑ 207 TGG ﻪـﺑ CGG ﻲــﻣ ﻦﻴــﻧژرآﻪــﻨﻴﻣآﺪﻴــﺳانآﻪــﺠﻴﺘﻧردﻪــﻛدﻮــﺷ نﺎــﻓﻮﺘﭙﻳﺮﺗﻦﻳﺰﮕﻳﺎــﺟ ﻲــﻣ دﻮــﺷ . عﻮــﻧزانﻮﻴــﺳﺎﺗﻮﻣ Missence mutation ــﺟوهدﻮــﺑ ﻪــﻨﻴﻣآﺪﻴــﺳاﻲﻨﻳﺰﮕﻳﺎ نﺎﻤﺘﺧﺎـﺳردلﻼﺘـﺧاﺚـﻋﺎﺑنﺎـﻓﻮﺘﭙﻳﺮﺗيﺎﺟﻪﺑ ﻦﻴﻧژرآ هﺮـﻴﺠﻧزﻲﻳﺎﻀـﻓ GP Ibα دﺮـﻜﻠﻤﻋﻪـﻛيﻮـﺤﻧﻪـﺑهﺪـﺷ ﻞﺘﺨﻣارنآيراﺪﻳﺎﭘﻻﺎﻤﺘﺣاو ﻲـﻣ ﺪـﻨﻛ . نﺎـﻓﻮﺘﭙﻴﻳﺮﺗ 207 ﻲﻟاﻮﺗويﺪﻴﻔﻟﻮﺳيدپﻮﻟرﺎﺘﺧﺎﺳﻞﺧادرد يﺎﻫ زاﻲﻨﻏ ﺎـﺑﻪـﻘﻄﻨﻣﻦـﻳاردوﻪﺘـﺷادراﺮﻗﻦﻴﺳﻮﻟ 7 ﻪـﻨﻴﻣآﺪﻴـﺳا ﻪﻛنآهﺪﻨﻨﻛﻪﻃﺎﺣا 4 نآيﺎﺗ ﺎﻫ ﻲﻟاﻮـﺗﻦﻳﺮـﺧآﻞـﺧادرد وﻦﻴـــﺳﻮﻟزاﻲـــﻨﻏ 3 نآيﺎـــﺗ ﺎـــﻫ يدپﻮـــﻟنوردرد طﺎـﺒﺗراﻦـﻳاﻪﻛهدﻮﺑطﺎﺒﺗراردﺪﻧا ﻪﺘﻓﺮﮔراﺮﻗيﺪﻴﻔﻟﻮﺳ نﺎﻓﻮﺘﭙﻳﺮﺗﻲﻨﻳﺰﮕﻳﺎﺟوهدﻮﺑﻚﻴﺑﻮﻓورﺪﻴﻫعﻮﻧزا 207 ﺎـﺑ ﻪﻨﻴﻣآﺪﻴﺳاﺮﻳﺎﺳ يﺎﻫ ندرﻮـﺧﻢﻬﺑﺚﻋﺎﺑبﻮﻓورﺪﻴﻫﺮﻴﻏ هﺪﺷطﺎﺒﺗراﻦﻳا ﺮـﻴﻏﻲﮔدرﻮـﺧﺎـﺗهﺮـﻴﺠﻧزﻪـﺠﻴﺘﻧردو يراﺪـﻳﺎﭘﺎﻧوهﺮـﻴﺠﻧزﻪﻳﺰﺠﺗﻊﻳﺮﺴﺗﻪﺑوهدﺮﻛاﺪﻴﭘﻲﻌﻴﺒﻃ ﺮﺠﻨﻣنآ ﻲﻣ دﻮﺷ . ) 7 ( ﻚﻳنﺎﻓﻮﺘﭙﻳﺮﺗ هﺪﺷﻆﻔﺣﻪﻨﻴﻣآﺪﻴﺳا ) Conserved ( هﺪـﻨﻫدﻞﻴﻜﺸﺗهﺮﻴﺠﻧزرﺎﻬﭼﺮﻫ ردوهدﻮﺑ هﺪﻳد ﺲﻜﻠﭙﻤﻛ ﻲﻣ دﻮﺷ . ﻦﻴﺌﺗوﺮـﭘﺮﻳﺎﺳردهوﻼﻋﻪﺑ يﺎـﻫ ﻲــﻨﻏيﺎــﻫراﺮﻜﺗيارادهداﻮﻧﺎــﺧ ﻦــﻳاردﻢــﻫﻦﻴــﺳﻮﻟزا ﻪﻨﻴﻣآﺪﻴـﺳا ﺮﻳﺎﺳﺎﻳنﺎﻓﻮﺘﭙﻳﺮﺗﺖﻴﻌﻗﻮﻣ يﺎـﻫ بﻮﻓورﺪـﻴﻫ راﺮـﻗﻦﻴﻧﻻآﻞﻴﻨﻓﺎﻳﻦﻴﺳﻮﻟﻞﺜﻣ ﻲـﻣ ﺮـﻣاﻦﻴـﻤﻫ ﻪـﻛدﺮﻴﮔ دﺮـﻜﻠﻤﻋورﺎﺘﺧﺎـﺳﻆـﻔﺣردارﻪـﻨﻴﻣآﺪﻴـﺳاﻦـﻳاﺖﻴﻤﻫا ﻦﻴﺌﺗوﺮﭘ يﺎـﻫ ﻲﻟاﻮـﺗياراد يﺎـﻫ نﺎﺸـﻧﻦﻴـﺳﻮﻟزاﻲـﻨﻏ ﻲﻣ حﺮﻄﻣوﺪﻫد ﻲﻣ ﻪـﺑﻲﺘﻴﺴـﻴﺑﻮﻓورﺪﻴﻫﺮـﻴﻴﻐﺗﻪـﻛﺪـﻨﻛ د ﻪﻨﻴﻣآﺪﻴﺳاﻲﻨﻳﺰﮕﻳﺎﺟﻞﻴﻟ يﺎﻫ ﻪـﻘﻄﻨﻣﻦـﻳاردﻞﻴﻓورﺪﻴﻫ نآدﺮــﻜﻠﻤﻋردلﻼﺘــﺧاوهﺮــﻴﺠﻧزيراﺪــﻳﺎﭘﺎﻧﺚــﻋﺎﺑ ﻲﻣ ﺪﻧﻮﺷ . ) 7 ( شﺮـﺑﻞـﺤﻣﻚـﻳدﺎﺠﻳاﺚﻋﺎﺑنﻮﻴﺳﺎﺗﻮﻣﻦﻳا ياﺮﺑ 1 Msp ﻲﻣ ﻪـﻛدﻮـﺷ ﻲـﻣ شورﻲـﺣاﺮﻃﻪـﻳﺎﭘناﻮـﺗ RFLP يرﺎـﻤﻴﺑﺺﻴﺨﺸـﺗياﺮﺑنآزاودﻮﺷهدادراﺮﻗ ﻗﺎﻧﻦﻴﻴﻌﺗوﻪﺘﺴﺑاوداﺮﻓارد ﺪـﻟﻮﺗزاﻞـﺒﻗﺺﻴﺨﺸﺗوﻦﻴﻠ دﺮﻛهدﺎﻔﺘﺳارﺎﻤﻴﺑنﺎﮕﺘﺴﺑرد . نﻮﻴﺳﺎﺗﻮﻣ A 710 G ﺶـﻬﺟﻚـﻳﻪﻛ چﻮـﭘ ﺚـﻋﺎﺑ،ﺖـﺳا ﺮﻴﻴﻐﺗ TGG نﺎﻓﻮﺘﭙﻳﺮﺗنوﺪﻛ 207 ﺮـﮔﻪـﻤﺗﺎﺧنوﺪـﻛﻪـﺑ TAG ﻲﻣ هﺮـﻴﺠﻧزنﻮﻴﺳﺎﺗﻮﻣﻦﻳاﻪﺠﻴﺘﻧردﻪﻛدﻮﺷ GP Ibα ﺰﺘﻨﺳﻲﺼﻗﺎﻧ ﻲـﻣ ﻪـﻛدﻮـﺷ 206 لﻮـﻃﻪـﻨﻴﻣآﺪﻴـﺳا ﻲﻟاﻮــﺗﺪــﻗﺎﻓوﻪﺘــﺷاد يﺎــﻫ ﻼﭘﻮﺘﻴﺳ ﻞــﺧادﻲﻟاﻮــﺗ،ﻲﻤــﺳ ﺪﻴﺘﭙﭘﻮﻜﻴﻠﮔوﺮﻛﺎﻣﻦﻣودوﻲﻳﺎﺸﻏ ﻲﻣ ﺪﺷﺎﺑ . هﺮـﻴﺠﻧزﻦـﻳا هﺮـﻴﺠﻧزﺮﻳﺎـﺳﺎﺑطﺎﺒﺗرادﺎﺠﻳاﻪﺑردﺎﻗﺺﻗﺎﻧ ﺎـﻫ ،هدﻮـﺒﻧ ﻞﺘﺨﻣﺖﻛﻼﭘﺎﺸﻏﺢﻄﺳردنآيﺮﻴﮔراﺮﻗ ﻲﻣ دﻮﺷ . ﺮﺑﺎﻨﺑ رﺎﻈﺘﻧاﻦﻳا ﻲﻣ دﻮـﺷﻪـﻳﺰﺠﺗﺰﺘﻨـﺳزاﺲـﭘهﺮـﻴﺠﻧزدور . ﺘﻬﺑكردياﺮﺑﻲﻟﻮﻠﺳنﺎﻴﺑ وﻲﻳﺎﻴﻤﻴﺷﻮﻴﺑتﺎﻌﻟﺎﻄﻣ ﻦﻳاﺮ ﺖﺳامزﻻنﻮﻴﺳﺎﺗﻮﻣ . ﻚـﻳفﺬـﺣﺚـﻋﺎﺑنﻮﻴـﺳﺎﺗﻮﻣﻦﻳا ﻢﻳﺰﻧآياﺮﺑشﺮﺑﻞﺤﻣ EcorII ﻲﻣ ﻪﻛدﻮﺷ ﻲﻣ ﻪﻳﺎﭘﺪﻧاﻮﺗ شور ﻲﺣاﺮﻃ RFLP ﺮﻳﺎـﺳردنﻮﻴﺳﺎﺗﻮﻣيﺮﻴﮕﻴﭘياﺮﺑ دﺮﻴﮔراﺮﻗﻦﻴﻠﻗﺎﻧﻦﻴﻴﻌﺗوﻪﺘﺴﺑاوداﺮﻓا . نﻮﻴﺳﺎﺗﻮﻣ T 1759 C نﻮﻴﺳﺎﺗﻮﻣعﻮﻧﻚﻳﻢﻫ چﻮﭘ ﺖﺳا ﺮﻴﻴﻐﺗﺚﻋﺎﺑﻪﻛ CGA ﻦﻴﻧژرآنوﺪﻛ 557 ﻪﺑ TGA نوﺪﻛ ﻲﻣﺮﮔﻪﻤﺗﺎﺧ هﺮـﻴﺠﻧزﻲﻤـﺳﻼﭘﻮﺘﻴﺳﻲﻟاﻮﺗودﻮﺷ GP Ibα ﻲﻣراﺮﻗﺮﻴﺛﺎﺗﺖﺤﺗار ﺪﻫد . نﻮﻴﺳﺎﺗﻮﻣﻦﻳاﻪﺠﻴﺘﻧرد ﻲﻣﺰﺘﻨﺳﻲﺼﻗﺎﻧيﺪﻴﺘﭙﭘ ﻲﻠﭘهﺮﻴﺠﻧز ﻪﻛدﻮﺷ 556 ﺪﻴﺳا ﻦـﻣودﺪـﺟاو ﺺﻗﺎـﻧهﺮـﻴﺠﻧزﻦﻳادرادلﻮﻃ ﻪﻨﻴﻣآ يﺎـﻫ ﻲﻟاﻮـﺗزاﻲﺸـﺨﺑﺎﻣاهدﻮﺑﻲﻳﺎﺸﻏﻞﺧادوﻲﻟﻮﻠﺳجرﺎﺧ ﻴﺳ ﺖﺳاهﺪﺷفﺬﺣنآﻲﻤﺳﻼﭘﻮﺘ . هﺪـﺷفﺬـﺣﺖﻤﺴـﻗ ﻦﻴﺌﺗوﺮﭘونﺎﻣﻼﻴﻓﻪﺑ لﺎﺼﺗاياﺮﺑمزﻻ ﻲﻟاﻮﺗ يﺎﻫ ζ 3 -3 -14 ﻲﻣ ﺪـﺷﺎﺑ . ﻲـﻣنﺎﺸـﻧﻒـﻠﺘﺨﻣ تﺎـﻌﻟﺎﻄﻣ ﻪـﻛﺪـﻫد

(7)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار هﺮﻴﺠﻧزﻲﻤﺳﻼﭘﻮﺘﻴﺳﻲﻟاﻮﺗطﺎﺒﺗرا هﺪـﻨﻫدﻞﻴﻜﺸـﺗيﺎـﻫ ﻦﻴﺌﺗوﺮﭘﺎﺑﺲﻜﻠﭙﻤﻛ يﺎﻫ ζ 3 -3 -14 ﻲﮔﺪـﻨﻨﻛ ﻢﻴﻈﻨﺗﺶﻘﻧ ﻧرادهﺪﻬﻋﺮﺑ ارﺲﻜﻠﭙﻤﻛدﺮﻜﻠﻤﻋ ﺪ . ﻦـﻳاطﺎﺒﺗراهوﻼﻋﻪﺑ ﻲﻟاﻮﺗ ﻲﻟﻮﻠـﺳ نﻮﺘﻠﻜـﺳاﻮﺘﻴﺳﻆـﻔﺣﺚـﻋﺎﺑنﺎﻣﻼﻴﻓ ﺎﺑﺎﻫ ﻲﻣﺲﻜﻠﭙﻤﻛﺖﻴﻟﺎﻌﻓﻢﻴﻈﻨﺗو دﻮﺷ . ) 12 -8 ( ﻪﻛﻦﻳاﻪﺑﻪﺟﻮﺗﺎﺑ ﻲﻟاﻮﺗرﺎﻤﻴﺑﻦﻳارد C هﺮـﻴﺠﻧزلﺎـﻨﻴﻣﺮﺗ GP Ibα يوﺎـﺣ ﻞﺤﻣ ﻦﻴﺌﺗوﺮﭘياﺮﺑلﺎﺼﺗايﺎﻫ ζ 3 -3 -14 نﺎﻣﻼﻴﻓو A ، ﺖـﺳاهﺪـﺷفﺬﺣ ، ـﺳاﻮﺘﻴﺳﺎـﺑﺲﻜﻠﭙـﻤﻛطﺎـﺒﺗرا نﻮﺘﻠﻜ ﺑﻲﻟﻮﻠﺳ ﻪ ﻲﻣ ﻢﻫ درﻮﺧ مﺎـﻴﭘلﺎـﻘﺘﻧاﺪـﻧورهوﻼـﻋﻪـﺑو مﺎﻴﭘيراﺮﻗﺮﺑﻪﺑ ردﺎﻗﺲﻜﻠﭙﻤﻛوهﺪﺷﻞﺘﺨﻣ ﻲـﺷﺎﻧ يﺎﻫ ﺪﻧﺎﮕﻴﻟﻪﺑ ﻲﻟﻮﻠﺳجرﺎﺧﻦﻣودلﺎﺼﺗازا ﻲﻤﻧﺎﻫ ﺪﺷﺎﺑ . ﻦﻳا ﻲﻣنﺎﺸﻧﺐﻟﺎﻄﻣ ردلﻼﺘـﺧاﺚـﻋﺎﺑنﻮﻴـﺳﺎﺗﻮﻣﻪـﻛﺪـﻫد وهﺪﺷﺲﻜﻠﭙﻤﻛدﺮﻜﻠﻤﻋ طﺎﺒﺗراومﺎﻴﭘلﺎﻘﺘﻧاﻊﻳﺎﻗوﺪﻧور ﺎــﺑنآ ﻲــﻣﻞــﺘﺨﻣارﻲﻟﻮﻠــﺳنﻮﺘﻠﻜــﺳاﻮﺘﻴﺳ ﺪــﻨﻛ . ﻦــﻳا ﻢﻳﺰـﻧآياﺮـﺑشﺮـﺑﻞـﺤﻣﻚـﻳفﺬـﺣﺚـﻋﺎﺑنﻮﻴﺳﺎﺗﻮﻣ هﺪﻨﻨﻛدوﺪﺤﻣ 1 Taq ﻲﻣ ﻲـﻣﻪﻛدﻮﺷ ﻲـﺣاﺮﻃﻪـﻳﺎﭘﺪـﻧاﻮﺗ شور RFLP درﻮــﻣﻦﻴﻠﻗﺎــﻧﻦﻴــﻴﻌﺗردودﺮــﻴﮔراﺮــﻗ دﺮﻴﮔراﺮﻗهدﺎﻔﺘﺳا . فﺬﺣ 20 ﺖـﻴﻌﻗﻮﻣرد ﺪﻴﺗﻮﺌﻠﻛﻮﻧ 1819 -1800 ﺚـﻋﺎﺑ ﻲﺼﻗﺎﻧهﺮﻴﺠﻧزﺰﺘﻨﺳ ﻲﻣ ﻮﺷ ﻪﻛد 597 ﻪﻨﻴﻣآﺪﻴﺳا لﻮـﻃ ﺰﺘﻨﺳزاﺲﭘهﺮﻴﺠﻧزوﻪﺘﺷاد 29 ﻪـﻨﻴﻣآﺪﻴﺳا ﺪـﻳﺪﺟ ﻪـﺑ نﺎﻳﺎﭘ ﻲﻣ ﺪﺳر . ﺺﻗﺎﻧهﺮﻴﺠﻧزﻦﻳا 13 هﺎﺗﻮﻛﻪﻨﻴﻣآﺪﻴﺳا ﺮﺗ ﻲﻟاﻮﺗوهدﻮﺑﻲﻠﺻاهﺮﻴﺠﻧززا C لﻮﻃﻪﺑنآلﺎﻨﻴﻣﺮﺗ 29 ﻲﻟاﻮﺗزاﻪﻨﻴﻣآﺪﻴﺳا C توﺎـﻔﺘﻣﻢﻟﺎـﺳهﺮـﻴﺠﻧزلﺎـﻨﻴﻣﺮﺗ ﺖﺳا . تﺎﻌﻟﺎﻄﻣ Invitro هدادنﺎﺸﻧ فﺬـﺣﻪـﻛﺖﺳاهﺪﺷ C هﺮﻴﺠﻧزلﺎﻨﻴﻣﺮﺗ GP Ibα يوﺎﺣﻪﻛ 609 Ser ﺖﺳا ، ﻞﺤﻣ ﻦﻴﺌﺗوﺮـﭘياﺮـﺑ ﻻﺎـﺑﻞﻳﺎﻤﺗﺎﺑلﺎﺼﺗا ζ 3 -3 -14 وهدﻮـﺑ ﻲﻟاﻮﺗﻪﺑﻦﻴﺌﺗوﺮﭘﻦﻳالﺎﺼﺗا C هﺮﻴﺠﻧزلﺎﻨﻴﻣﺮﺗ GP Ibα ﺖﺳاﺲﻜﻠﭙﻤﻛدﺮﻜﻠﻤﻋهﺪﻨﻨﻛﻢﻴﻈﻨﺗ . ) 12 و 13 ( رﺎﻈﺘﻧاﻦﻳاﺮﺑﺎﻨﺑ ﻲــﻣ ﻦﻴﺌﺗوﺮــﭘطﺎــﺒﺗراﺶــﻬﺟﻦــﻳادور يﺎــﻫ ζ 3 -3 -14 و وهدﺮﻛﻞﺘﺨﻣارﺲﻜﻠﭙﻤﻛﻲﻤﺳﻼﭘﻮﺘﻴﺳﻦﻣودﻪﺑنﺎﻣﻼﻴﻓ ﺪـﻫﺪﺑﺖـﺳدزااردﻮـﺧدﺮـﻜﻠﻤﻋﺲﻜﻠﭙﻤﻛﻪﺠﻴﺘﻧرد . ﻪـﺑ نﺪــﺷهدوﺰــﻓاهوﻼــﻋ 29 ﻲﻟاﻮــﺗردﺪــﻳﺪﺟﻪــﻨﻴﻣآﺪﻴــﺳا هﺮــﻴﺠﻧزﻲﻤــﺳﻼﭘﻮﺘﻴﺳ GP Ibα ﺚــﻋﺎﺑﺖــﺳاﻦــﻜﻤﻣ، ﻲـﺳرﺮﺑﻪـﺑزﺎﻴﻧﻪﻛدﻮﺷ هﺮﻴﺠﻧزﻲﺘﻴﺴﻴﺑﻮﻓورﺪﻴﻫﺮﻴﻴﻐﺗ ﺰﻴﻣﻦﻴﻴﻌﺗوﻲﻳﺎﻴﻤﻴﺷﻮﻴﺑ هدﺎﻔﺘـﺳاﺎـﺑﻲﺘﻴﺴﻴﺑﻮﻓورﺪﻴﻫنا زا مﺮﻧ راﺰﻓا ﻞـﺜﻣدﻮـﺟﻮﻣيﺎﻫ Genepro software دراد . نﻮﻴـﺳﺎﺗﻮﻣﺮﻳﺎﺳﻪﺑﻪﻴﺒﺷ يﺎـﻫ ﻦـﻳاردهﺪـﺷﻲﻳﺎـﺳﺎﻨﺷ ﻲـﻠﺒﻗتﺎـﻌﻟﺎﻄﻣردوهدﻮـﺑﺪﻳﺪﺟنﻮﻴﺳﺎﺗﻮﻣﻦﻳا،ﻪﻌﻟﺎﻄﻣ ﺖﺳاهﺪﺸﻧشراﺰﮔﺎﻴﻧدﻒﻠﺘﺨﻣطﺎﻘﻧرد . ﻦـﻳاهوﻼـﻋﻪـﺑ هاﺮــﻤﻫﻪــﺑنﻮﻴــﺳﺎﺗﻮﻣ T 1759 C ﻮﻴــﺳﺎﺗﻮﻣﻦﻴــﻟوا ن يﺎــﻫ هﺮـﻴﺠﻧزﻲﻤـﺳﻼﭘﻮﺘﻴﺳﻦـﻣودردهﺪﺷشراﺰﮔ GP Ibα ﻲﻣ ﻦﻴـﻟوانﻮﻴـﺳﺎﺗﻮﻣﻦﻳارﻮﻃﻦﻴﻤﻫوﺪﻨﺷﺎﺑ Deletion mutation ﻲﻤــﺳﻼﭘﻮﺘﻴﺳﻦــﻣودرد GP Ibα ﻲــﻣ ﺪــﺷﺎﺑ . ﻢﻳﺰـﻧآ شﺮـﺑيﻮـﮕﻟاﺮﻴﻴﻐﺗ ﺚﻋﺎﺑنﻮﻴﺳﺎﺗﻮﻣ ﻦﻳارﻮﻀﺣ هﺪﻨﻨﻛدوﺪﺤﻣ 1 Bsiy ﻲﻣ ﻪﻛدﻮﺷ ﻲـﻣ ﻲـﺣاﺮﻃﻪـﻳﺎﭘﺪـﻧاﻮﺗ شور RFLP ﻲﺑﺎﻳدرياﺮﺑ نﺎﮕﺘﺴـﺑردنﻮﻴـﺳﺎﺗﻮﻣﻦﻳا دﺮﻴﮔراﺮﻗرﺎﻤﻴﺑ . نﻮﻴـﺳﺎﺗﻮﻣﺶﻘﻧدرﻮﻣردﺎﻣتﺎﻋﻼﻃازايرﺎﻴﺴﺑ ﺎـﻫ درﺎـﻧﺮﺑنارﺎﻤﻴﺑ ﻪﻌﻟﺎﻄﻣزا،ﺲﻜﻠﭙﻤﻛدﺮﻜﻠﻤﻋﺮﺑ ﺮﻴﻟﻮـﺳ ﺖــﺳاهﺪــﺷﻞــﺻﺎﺣ . ﻦــﻣودردنﻮــﻨﻛﺎــﺗﻪــﻛﺎــﺟنآزا تﺎـﻋﻼﻃا،دﻮـﺑهﺪﺸـﻧشراﺰﮔﻲﻧﻮﻴﺳﺎﺗﻮﻣﻲﻤﺳﻼﭘﻮﺘﻴﺳ نﻮﻴـﺳﺎﺗﻮﻣﻦـﻳاﺶـﻘﻧﻪـﺑﻊـﺟارﻲﺻﺎﺧ ﺎـﻫ ا نارﺎـﻤﻴﺑز ﺖــﺳاهﺪــﻣﺎﻴﻧﺖــﺳﺪﺑﺮﻴﻟﻮــﺳدرﺎــﻧﺮﺑ . ﺮــﺘﻬﺑكردياﺮــﺑ نﻮﻴــﺳﺎﺗﻮﻣﻦــﻳاﺶــﻘﻧ ﺎــﻫ ﺲﻜﻠﭙــﻤﻛنﺎــﻴﺑوﻪــﺿﺮﻋرد يروﺮـﺿ ﻲﻟﻮﻠﺳنﺎﻴﺑوﻲﻳﺎﻴﻤﻴﺷﻮﻴﺑتﺎﻌﻟﺎﻄﻣ ﻲـﻣ ﺪـﺷﺎﺑ ﺖﻳدوﺪﺤﻣزاﻪﻛ يﺎـﻫ ﺖـﺳاﺪـﻴﻣاوهدﻮـﺑﺶﻫوﮋـﭘﻦـﻳا ﻢﻫاﺮﻓرﺎﻛﻦﻳاياﺮﺑارﻲﺒﺳﺎﻨﻣﺮﺘﺴﺑهﺪﺷمﺎﺠﻧاﺶﻫوﮋﭘ دروآ . ﻴﺑدرﻮﻣردﻦﻴﻨﭽﻤﻫ هﺮﻴﺠﻧزنژ ردﻪﻛﻲﻧارﺎﻤ GP Ibα ﺮﻳﺎـــﺳنژﻲـــﺳرﺮﺑﺪـــﻧدﻮﺒﻧنﻮﻴـــﺳﺎﺗﻮﻣياراد هﺮــﻴﺠﻧز ﺎــﻫ نآﻪــﺑﻪــﻌﻟﺎﻄﻣﻦــﻳاردﻪــﻛهدﻮــﺑيروﺮــﺿ ﺖﺳاهﺪﺸﻧﻪﺘﺧادﺮﭘ . هﺮـﻴﺠﻧزنژﻲﻟﻮﻜﻟﻮﻣﺺﻳﺎﻘﻧﻪﻌﻟﺎﻄﻣﻦﻳارد GP Ibα وﺖـﻓﺮﮔراﺮﻗﻪﻌﻟﺎﻄﻣدرﻮﻣ 5 ﻪـﻛﺪـﻳﺪﺟ نﻮﻴـﺳﺎﺗﻮﻣ رد ﺎـﺳﺎﻨﺷ،دﻮـﺑهﺪﺸـﻧﻲﻳﺎﺳﺎﻨﺷتﺎﻌﻟﺎﻄﻣﺮﻳﺎﺳ ﺪـﻳدﺮﮔﻲﻳ . نﻮﻴـﺳﺎﺗﻮﻣزاماﺪـﻛﺮـﻫياﺮﺑ ﺎـﻫ RFLP ) (Restriction

(8)

ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻦﻴﺌﺗوﺮﭘﻮﻜﻴﻠﮔ نژ رد Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار

Fragment Length Polymorphism

ﺪﺷمﺎﺠﻧاوﻲﺣاﺮﻃ . ﺺﻗاﻮــﻧﻲﻳﺎــﺳﺎﻨﺷونژﻦــﻳاﻪــﻌﻟﺎﻄﻣشورﻲــﻓﺮﻌﻣ نآﻲﻟﻮﻜﻟﻮﻣ ﻲﻣ ﺺﻴﺨﺸﺗزﺎﻏآﺪﻧاﻮﺗ يﺎﻫ ﻦﻴﻴﻌﺗ،ﻲﻟﻮﻜﻟﻮﻣ يرﺎـﻤﻴﺑتﺎـﻋﻼﻃاﻚـﻧﺎﺑﻞـﻴﻤﻜﺗﻪـﺑﻚـﻤﻛوﻦﻴﻠﻗﺎﻧ يﺎـﻫ ﻧهﺪﻨﻫديﺰﻳﺮﻧﻮﺧ ﺪﺷﺎﺑرﻮﺸﻛردردﺎ .

ﺮﻜﺸﺗ

و

ﺮﻳﺪﻘﺗ

ﻦﻳا ﻖﻴﻘﺤﺗ ﺎﺑ هدﺎﻔﺘﺳا زا ﺖﻳﺎﻤﺣ ﻲﻟﺎﻣ هﺎﮕﺸـﻧاد مﻮـﻠﻋ ﻲﻜﺷﺰﭘ و تﺎﻣﺪﺧ ﻲﺘﺷاﺪﻬﺑ ﻲﻧﺎﻣرد ناﺮﻳا رد ﺐـﻟﺎﻗ حﺮـﻃ ﺪﻛﻲﺗﺎﻘﻴﻘﺤﺗ 306 / پ مﺎﺠﻧا هﺪﻳدﺮﮔ ﺖﺳا ﻪﻛ ﻪﻠﻴـﺳوﻦﻳﺪـﺑ ﺐﺗاﺮﻣ ﺮﻳﺪﻘﺗ و ﺮﻜﺸﺗ ﻲﻣمﻼﻋانﺎﮔﺪﻨﺴﻳﻮﻧ ددﺮﮔ .

ﻊﺑﺎﻨﻣ

ﺖﺳﺮﻬﻓ

1-Afshar-Kharghan V, Lo´pez JA. Bernard-Soulier

syndrome caused by a dinucleotide deletion and reading frame shift in the region encoding the glycoprotein Iba transmembrane domain. Blood. 1997; 90: 2634-643

2-Sachs UJH, Kroll H, Matzdorff AC, Berghofer H, Lopez JA, Santoso S. Bernard-Soulier syndrome due to the homozygous asn-45ser mutation in GPIX: an unexpected, frequent finding in Germany. Brit J Haemat. 2003; 123: 127-31

3-Rivera CE, Villagra J, Riordan M, Williams S, Lindstrom KJ, Rick ME. Identification of a new mutation in platelet glycoprotein IX in a patient with Bernard-Soulier syndrome. Br J Haem. 2001; 112: 105-8

4-Drouin J, Carson NL, Laneuville O. Compound heterozygosity for a novel nine-nucleotide deletion and the Asn45Ser missense mutation in the glycoprotein IX gene in a patient with Bernard-Soulier syndrome. AmJ Hematol. 2005; 78(1):41-48

5-Lopez JA, Andrews RK, Afshar-Kharghan V, Berndt M.C. Bernard-Soulier syndrome. Blood. 1998; 91: 4397-418

6-Provan A, Gribben J. Molecular hematology. 2nd

edition. Oxford: Blackwell science; 2005. p. 210-17 7-Rosenberg N, Lalezari S, Landau M, Shenkman B, Seligsohn U, Izraeli S. Trp207Gly in platelet glycoprotein Ibα is a novel mutation which disrupts the

connection between the leucine-rich repeat domain and the disulfide loop structure and causes Bernard-Soulier syndrome. J Thromb Haemost. 2007; 5: 378–86

8-Mangin P, David T, Lavaud V, Cranmer SL, Pikovski I, Jackson SP, et al. Identification of a novel 14-3-3ζ binding site within the cytoplasmic tail of platelet glycoprotein Ibζ. Blood. 2004; 104: 420-27

9-Feng S, Christodoulides N, Resendiz JC, Berndt MC, Kroll H. Cytoplasmic domains of GpIba and GpIbb regulate 14-3-3ζ binding to GpIb/IX/V. Blood. 2000; 95: 551-57

10-Berndt MC, Cranmer SL, Andrews RK. Filamin a interaction with GPIb_:the platelet shapes up. Blood. 2006; 107: 1745

11-Bodnar RJ, Xi X, Li Z, Berndt MC, Du X. Regulation of glycoprotein Ib-IX-von Willebrand factor interaction by cAMP-dependent protein kinase mediated phosphorylation at Ser 166 of glycoprotein Ib beta. J Biol Chem. 2002; 277: 47080-87

12-Bodnar RJ, Gu M, Li Z, Englund GD, Du X. The cytoplasmic domain of the platelet glycoprotein Ib_ is phosphorylated at serine609. J Biol Chem. 1999; 274: 33474-479

13-Dai K, Bodnar RJ, Berndt MC, Du X. A critical role for 14-3-3ζ protein in regulating the VWF binding function of platelet glycoprotein Ib-IX and its therapeutic implications. Blood. 2005; 106: 1975-981

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ﻲﻳﺎﺳﺎﻨﺷ 5 ﺪﻳﺪﺟ ﺶﻬﺟ ﻴﻠﮔ نژ رد ﻦﻴﺌﺗوﺮﭘﻮﻜ Ibα ﺖﻛﻼﭘ نارﺎﻤﻴﺑ رد درﺎﻧﺮﺑ -ناﺮﻳا ﺮﻴﻟﻮﺳ يﻮﻘﺗﻪﻟاﺪﺳاﺪﻴﺳ ﺎﻜﻤﻫو نار

Identification of Five Novel Mutations in Platelet GPIb

α

Gene

among Iranian Bernard-Soulier Patients

S.A. Taghavi, MScI * A. Kazemi, PhDII .Rastegar Lari, PhDIII

F. Ala, MDIV_{M. Rasoolzadegan, BSc}V

Abstract

Background & Aim: Bernard-Soulier syndrome (B.S.S) is a rare hereditary bleeding disorder due to

molecular defects of platelet GPIb–IX–V. The GPIb-IX-V complex is composed of four chains of GPIbα,

GPIbβ, GPIX and GPV. The largest chain of this complex is GPIbα and is responsible for binding to ligand

and most of identified mutations belong to this glycoprotein. The aim of this study was to identify the

molecular defects of GPIbα gene in Iranian Bernard-Soulier patients.

Patients and Method: Twelve Bernard-Soulier patients were selected from data base of bleeding

disorders in Comprehensive Clinic of Iranian Hemophilia Center. Diagnostic criteria for B.S.S were based

on phenotypic analysis such as platelet counts, inspection of peripheral blood smear and lack of response to restocetin agonist. Genomic DNA was isolated from blood leukocytes of the patients and their parents.

The entire amino acid coding region in exon 2 from GPIbα was divided into five overlapping fragments

and PCR amplification was done. Finally, sequence analysis of the coding regions that contain DNA heteroduplexes in CSGE gels was performed.

Results: Sequence analysis revealed five novel mutations in GPIbα. The mutations include ACCGGCT

deletion, GGA insertion in 419-425 position, missense mutations in T709C, G710A and C1759T, and the deletion of 20 nucleotides in 1800-1819 position. All five novel mutations were registered in International Gene Bank and for each mutation RFLP (restriction fragment length polymorphism) design was created.

Conclusion: In the present study, the molecular defects of GPIbα gene was investigated and five

novel mutations were identified among Iranian BSS patients.

Key Words: 1) Bernard-Soulier Syndrome 2) Glycoprotein Ibα

3) Sequence Analysis 4) Mutation

This research was financed by Iran University of Medical Sciences and Health Services. I) MSc in Hematology. Iran University of Medical Sciences and Health Services.Tehran, Iran.

II) Associate Professor of Hematology. Iran University of Medical Sciences and Health Services. Tehran, Iran.(*Corresponding Author)

III) Assistant Professor of Hematology. Iran University of Medical Sciences and Health Services. Tehran, Iran. IV) Subspecialist in Clinical Hematology. Comprehensive Clinic of Iranian Hemophilia center. Tehran, Iran. V) BSc in Chemistry. Comprehensive Clinic of Iranian Hemophilia center. Tehran, Iran.