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Evaluation of Myostatin Gene Polymorphism in Sanjabi Sheep by PCR-RFLP

Method

Article · January 2009

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6 authors, including:

Mohammad Reza Mohammadabadi Amin Baghizadeh

Shahid Bahonar University of Kerman Graduate University of advanced Technology
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Nahid Askari
Leeds City College
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 ‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‪ /‬ﺟﻠﺪ ‪ ١٩‬ﺷﻤﺎﺭﻩ ‪ /١‬ﺳﺎﻝ ‪١٣٨٨‬‬

‫ﺍﺭﺯﻳﺎﺑﯽ ﭼﻨﺪﺷﻜﻠﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﮋﺍﺩ ﺳﻨﺠﺎﺑﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺭﻭﺵ ‪PCR-RFLP‬‬

‫‪۲‬‬
‫ﺑﻬﺎﺭﻩ ﺻﻮﻓﯽ‪ ،۱‬ﻣﺤﻤﺪﺭﺿﺎ ﻣﺤﻤﺪﺁﺑﺎﺩﻱ‪ ،*۲‬ﮐﻤﺎﻝ ﺷﺠﺎﻋﻴﺎﻥ‪ ،۳‬ﺍﻣﻴﻦ ﺑﺎﻗﻲ ﺯﺍﺩﻩ‪ ،۴‬ﺳﻴﺮﻭﺱ ﻓﺮﺍﺳﺘﯽ‪ ،۵‬ﻧﺎﻫﻴﺪ ﻋﺴﮑﺮﯼ‪ ۱‬ﻭ ﺍﻣﻴﺪ ﺩﻳﺎﻧﯽ‬

‫ﺗﺎرﯾﺦ ﭘﺬﯾﺮش‪88/3/10 :‬‬

‫‪ -1‬داﻧﺸﺠﻮﯾﺎن ﮐﺎرﺷﻨﺎﺳﯽ ارﺷﺪ ﻋﻠﻮم داﻣﯽ‪ ،‬داﻧﺸﮕﺎه زاﺑﻞ‬
‫‪ -2‬ﺑﺨﺶ ﻋﻠﻮم داﻣﯽ‪ ،‬داﻧﺸﮑﺪه ﮐﺸﺎورزي‪ ،‬داﻧﺸﮕﺎه ﺷﻬﯿﺪ ﺑﺎﻫﻨﺮ ﮐﺮﻣﺎن‬
‫‪ -3‬ﺑﺨﺶ ﻋﻠﻮم داﻣﯽ‪ ،‬داﻧﺸﮑﺪه ﮐﺸﺎورزي‪ ،‬داﻧﺸﮕﺎه زاﺑﻞ‬
‫‪ -4‬ﻣﺮﮐﺰ ﺑﯿﻦ اﻟﻤﻠﻠﯽ ﻋﻠﻮم و ﺗﮑﻨﻮﻟﻮژي ﭘﯿﺸﺮﻓﺘﻪ و ﻋﻠﻮم ﻣﺤﯿﻄﯽ ﮐﺮﻣﺎن‬
‫‪ -5‬ﮐﺎرﺷﻨﺎس ارﺷﺪ ﻣﺮﮐﺰ ﺗﺤﻘﯿﻘﺎت ﻋﻠﻮم ﮐﺸﺎورزي و داﻣﯽ ﮐﺮﻣﺎﻧﺸﺎه‬
‫‪mmohammadabadi@yahoo.ca‬‬ ‫*ﻣﺴﺌﻮل ﻣﮑﺎﺗﺒﻪ‬

‫ﭼﮑﻴﺪﻩ‬
‫ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﻳﺎ ﻋﺎﻣﻞ ‪ ۸‬ﺭﺷﺪ ﻭ ﺗﻤﺎﻳﺰ )‪ (GDF8‬ﺑﻪ ﻋﻨﻮﺍﻥ ﻋﺎﻣﻞ ﺍﻳﺠﺎﺩ ﮐﻨﻨﺪﻩ ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﺷﻨﺎﺧﺘﻪ ﺷـﺪﻩ ﺍﺳـﺖ‪ ،‬ﮐـﻪ ﺩﺭ‬
‫ﺁﻥ ﻣﺠﻤﻮﻋﻪ ﺍﯼ ﺍﺯ ﺟﻬﺶ ﻫﺎﯼ ﻏﻴﺮ ﻓﻌﺎﻝ ﮐﻨﻨﺪﻩ ﮊﻥ ﺭﺥ ﻣﯽ ﺩﻫﻨﺪ‪ .‬ﺩﺭ ﮔﺎﻭ‪ ،‬ﺟﻬﺶ ﻫﺎﯼ ﻏﻴﺮ ﻓﻌـﺎﻝ ﻛﻨﻨـﺪﻩ ﺩﺭ ﺍﻳـﻦ ﮊﻥ ﻣﻨﺠـﺮ ﺑـﻪ‬
‫ﺗﻮﻟﻴﺪ ﻓﻨﻮﺗﻴﭗ ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﻣﻲ ﺷﻮﺩ‪ .‬ﭼﻨﺪ ﺷﮑﻠﯽ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﻴﺰ ﺑﺮﺭﺳﯽ ﺷﺪﻩ ﺍﺳـﺖ‪ .‬ﮔﻮﺳـﻔﻨﺪ ﺳـﻨﺠﺎﺑﯽ‬
‫ﻳﮑﯽ ﺍﺯ ﻧﮋﺍﺩﻫﺎﯼ ﺍﺻﻠﯽ ﮔﻮﺳﻔﻨﺪ ﺩﺭ ﺍﻳﺮﺍﻥ‪ ،‬ﺑﻪ ﻭﻳﮋﻩ ﺩﺭ ﺍﺳﺘﺎﻥ ﮐﺮﻣﺎﻧـﺸﺎﻩ ﺍﺳـﺖ ﮐـﻪ ﺗـﺎ ﮐﻨـﻮﻥ ﺑـﺮﺍﯼ ﺍﻳـﻦ ﺟﺎﻳﮕـﺎﻩ ﺑـﻪ ﻭﺳـﻴﻠﻪ‬
‫ﻧﺸﺎﻧﮕﺮﻫﺎﯼ ﻣﻮﻟﮑﻮﻟﯽ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺪﻩ ﺍﺳﺖ‪ .‬ﻫﺪﻑ ﺍﺯ ﺍﻧﺠﺎﻡ ﺍﻳﻦ ﻣﻄﺎﻟﻌﻪ ﺁﻧﺎﻟﻴﺰ ﻧﺎﺣﻴﻪ ﮐﺪ ﮐﻨﻨﺪﻩ ﺩﺭﺑﺮﮔﻴﺮﻧﺪﻩ ﺟﻬـﺶ ﻫـﺎﻳﯽ ﮐـﻪ ﺑـﻪ‬
‫ﻃﻮﺭ ﺑﺎﻟﻘﻮﻩ ﺑﻴﺎﻥ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺭﺍ ﺗﻐﻴﻴﺮ ﻣﯽ ﺩﻫﻨﺪ ﺑﻮﺩ‪ .‬ﺍﺯ ﺻﺪ ﻭ ﭘﻨﺠﺎﻩ ﺭﺃﺱ ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﯽ ﻧﻤﻮﻧﻪ ﻫﺎﯼ ﺧﻮﻥ ﮔﺮﻓﺘـﻪ ﺷـﺪ‬
‫ﻭ ‪ DNA‬ﺍﺳﺘﺨﺮﺍﺝ ﺷﺪﻩ ﺑﺮﺍﯼ ﺗﮑﺜﻴﺮ ﻗﻄﻌﻪ ‪ ۳۳۷‬ﺟﻔﺖ ﺑﺎﺯﯼ ﺍﺳﺘﻔﺎﺩﻩ ﺷﺪ‪ .‬ﭼﻨﺪ ﺷﮑﻠﯽ ﻃﻮﻝ ﻗﻄﻌﺎﺕ ﺑﺮﺷﯽ ﻣﺤﺼﻮﻻﺕ ‪ PCR‬ﺑﺎ‬
‫ﺍﻓـﺰﻭﺩﻥ ﺁﻧــﺰﻳﻢ ﺑﺮﺷــﯽ ‪ HaeIII‬ﺑــﻪ ﻭﺍﮐــﻨﺶ ‪ PCR‬ﮐﺎﻣــﻞ ﺍﺟـﺮﺍ ﺷــﺪ‪ .‬ﮊﻧﻮﺗﻴــﭗ ﻫــﺎﯼ ‪ PCR-RFLP‬ﺁﻧــﺎﻟﻴﺰ ﺷــﺪﻧﺪ‪ .‬ﻓﺮﺍﻭﺍﻧــﻲ‬
‫ﮊﻧﻮﺗﻴﭗ ﻫﺎﻱ ‪ Mm، MM‬ﻭ‪ mm‬ﺑﻪ ﺗﺮﺗﻴﺐ ‪ %۱/۳۳ ،%۲‬ﻭ ‪ %۹۶/۷۶‬ﺗﺸﺨﻴﺺ ﺩﺍﺩﻩ ﺷﺪﻧﺪ‪ .‬ﻓﺮﺍﻭﺍﻧﻲ ﺁﻟﻠﻲ ﻧﻴﺰ ﺑﺮﺍﻱ ﺁﻟـﻞ ﻫـﺎﻱ ‪M‬‬
‫ﻭ ‪ m‬ﺑﻪ ﺗﺮﺗﻴﺐ ‪ ۰/۰۳‬ﻭ ‪ ۰/۹۷‬ﺑﺮﺁﻭﺭﺩ ﮔﺮﺩﻳﺪ‪ .‬ﻣﻘﺎﻳﺴﻪ ﻓﺮﺍﻭﺍﻧﻲ ﺁﻟﻞ ‪) M‬ﺁﻟﻞ ﻣﻄﻠﻮﺏ( ﻣﺤﺎﺳﺒﻪ ﺷﺪﻩ ﺩﺭ ﮔﻮﺳﻔﻨﺪﻫﺎﻱ ﺳﻨﺠﺎﺑﻲ ﺑﺎ‬
‫ﺗﺤﻘﻴﻘﺎﺕ ﻣﺸﺎﺑﻪ ﺩﺭ ﻧﮋﺍﺩﻫﺎﻱ ﻣﺨﺘﻠﻒ ﺩﻧﻴﺎ ﻧﺸﺎﻥ ﺩﺍﺩ ﮐﻪ ﻓﺮﺍﻭﺍﻧﻲ ﺍﻳﻦ ﺁﻟﻞ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﻲ ﺩﺭ ﺳـﻄﺢ ﻣﻨﺎﺳـﺒﻲ ﻧﻤـﻲ ﺑﺎﺷـﺪ‪.‬‬
‫ﻋﻼﻭﻩ ﺑﺮ ﺍﻳﻦ‪ ،‬ﻣﺸﺨﺺ ﮔﺮﺩﻳﺪ ﮐﻪ ﺗﻌـﺎﺩﻝ ﻫـﺎﺭﺩﻱ‪ -‬ﻭﺍﻳﻨﺒـﺮﮒ ﺩﺭ ﺟﻤﻌﻴـﺖ ﻣـﻮﺭﺩ ﻣﻄﺎﻟﻌـﻪ ﺩﺭ ﺭﺍﺑﻄـﻪ ﺑـﺎ ﺍﻳـﻦ ﺟﺎﻳﮕـﺎﻩ ﺑﺮﻗـﺮﺍﺭ‬
‫ﻧﻤﻲ ﺑﺎﺷﺪ‪.‬‬

‫ﻭﺍﮊﻩﻫﺎﯼ ﮐﻠﻴﺪﻱ‪ :‬ﭼﻨﺪﺷﮑﻠﻲ‪ ،‬ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ‪ ،‬ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﻲ‪PCR-RFLP ،‬‬

 ١٣٨٨ ‫ ﺳﺎﻝ‬/١ ‫ ﺷﻤﺎﺭﻩ‬١٩ ‫ ﺟﻠﺪ‬/‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‬ ... ‫ ﻣﺤﻤﺪﺁﺑﺎﺩﯼ ﻭ‬،‫ﺻﻮﻓﯽ‬ ٨٢

Evaluation of Myostatin Gene Polymorphism in Sanjabi Sheep by PCR-RFLP

Method
B Soufy1, MR Mohammadabadi2, K Shojaeyan3, A Baghizadeh4, S Ferasaty5, N Askari1 and O Dayani2
1
MSc student of animal science, Faculty of Agriculture, Zabol University
2
Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman
3
Department Animal Science, Faculty of Agriculture, Zabpl University
4
International centre for science & high technology & environmental sciences
5
MSc, Agriculture and Animal Science Research Centre of Kermanshah
*Corresponding author: E-mail: mmohammadabadi@yahoo.ca

Abstract
Myostatin or growth and differentiation factor 8 (GDF8) has been identified as the factor causing a
phenotype known as double muscling in which a series of mutations render the gene inactive, and
therefore, unable to regulate muscle fibre deposition. Dysfunction of myostatin gene has been
reported in mammals. In bovine the loss of this gene activity has been associated to double-muscled
phenotype observed in some European cattle breeds. Myostatin gene polymorphism has also been
studied in sheep. Sanjabi sheep breed is one of the major sheep breeds in Iran, especially in
Kermanshah province that until now has not been studied at this locus by molecular markers. Due
to the role of myostatin gene in muscle development, the objective of this study was to analyze a
coding region containing mutations which potentially altering the myostatin gene expression. DNA
from blood samples of one hundred fifty Sanjabi sheep was extracted and used to amplify a 337-bp
fragment in myostatin gene. Restriction fragment length polymorphism (RFLP) of the PCR product
was performed by addition of HaeIII enzyme to the completed PCR reaction. PCR-RFLP genotypes
were analyzed. Genotype frequencies of MM, Mm and mm were detected as 2.00%, 1.33% and
96.70%, respectively. Allele frequencies were estimated for M and m alleles as 3.00% and 97.00%,
respectively. The data from this study indicated that the Sanjabi sheep was polymorphic for
myostatin gene though it was not at Hardy–Weinberg equilibrium.

Keyword: PCR-RFLP, Polymorphism, Myostatin gene, Sanjabi sheep

 ‫‪٨٣‬‬
‫‪ DM ،N‬ﻳــﺎ ‪ C ،dm‬ﻳــﺎ ‪ c ،N‬ﻳــﺎ ‪ A ،n‬ﻳــﺎ ‪ a‬ﻭ ‪ mh‬ﻳــﺎ ‪.+‬‬
‫ﻓﻨﻮﺗﻴﭗ ‪ DM‬ﺑﺎ ﻫـﺎﻳﭙﺮﺗﺮﻭﻓﯽ ﻣﺎﻫﻴﭽـﻪ ﻫـﺎ ﻣـﺸﺨﺺ ﺷـﺪﻩ‬
‫ﺍﺳﺖ‪ .‬ﺩﺭ ﺍﻳﻦ ﺣﻴﻮﺍﻧﺎﺕ ﺍﻧﺪﺍﻡ ﻫﺎﯼ ﺧﻠﻔﯽ ﺑﻪ ﺻـﻮﺭﺕ ﮔـﺮﺩ ﻭ‬
‫ﺑﺮﺟﺴﺘﻪ ﻫﺴﺘﻨﺪ ﻭ ﻣﺎﻫﻴﭽﻪ ﻫﺎ ﺣﺎﻟﺖ ﺑﻴﺮﻭﻥ ﺯﺩﮔﯽ ﺩﺍﺭﻧـﺪ ﻭ‬
‫ﺧﻄﻮﻁ ﺁﺷﻜﺎﺭﯼ ﺩﺭ ﺯﻳﺮ ﭘﻮﺳﺖ ﻗﺎﺑﻞ ﺩﻳﺪﻥ ﺍﺳﺖ‪ .‬ﺍﺯ ﺩﻳﮕـﺮ‬
‫ﻣﺸﺨﺼﻪ ﻫﺎﯼ ﻓﻴﺰﻳﻜﯽ ﺁﺷﻜﺎﺭ ﻣﯽ ﺗﻮﺍﻥ ﺑﻪ ﺍﺳﺘﺨﻮﺍﻥ ﻫـﺎﯼ‬
‫ﻇﺮﻳﻒ‪ ،‬ﺩﺳﺘﮕﺎﻩ ﺗﻨﺎﺳﻠﯽ ﻧﺎﺑﺎﻟﻎ ﻭ ﺯﺑـﺎﻥ ﺑـﺰﺭﮒ ﺩﺭ ﮔﻮﺳـﺎﻟﻪ‬
‫ﻫﺎﯼ ﺗﺎﺯﻩ ﻣﺘﻮﻟﺪ ﺷـﺪﻩ ﺍﺷـﺎﺭﻩ ﻛـﺮﺩ‪ .‬ﺣﻴﻮﺍﻧـﺎﺕ ﺑـﺎ ﻓﻨﻮﺗﻴـﭗ‬
‫ﻣﺎﻫﻴﭽــﻪ ﻣــﻀﺎﻋﻒ ﺩﺍﺭﺍﯼ ﺍﺳــﺘﺨﻮﺍﻥ ﻭ ﭼﺮﺑــﯽ ﻛﻤﺘــﺮ ﻭ‬
‫ﻣﺎﻫﻴﭽﻪ ﺑﻴﺸﺘﺮﯼ ﻫﺴﺘﻨﺪ ﻭ ﻧـﺴﺒﺖ ﻗـﺴﻤﺖ ﻫـﺎﯼ ﺍﺭﺯﺷـﻤﻨﺪ‬
‫ﮔﻮﺷﺖ ﺑﻴﺸﺘﺮ ﺍﺳﺖ‪ .‬ﺍﺯ ﻣﻌﺎﻳﺐ ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﻣﯽ ﺗـﻮﺍﻥ‬
‫ﺑـﻪ ﻛـﺎﻫﺶ ﺑـﺎﺭﻭﺭﯼ‪ ،‬ﻛــﺎﻫﺶ ﺑﻘـﺎﯼ ﮔﻮﺳـﺎﻟﻪ ﻫـﺎ‪ ،‬ﺍﻓــﺰﺍﻳﺶ‬
‫ﺣﺴﺎﺳﻴﺖ ﺑﻪ ﺍﺳﺘﺮﺱ ﻭ ﺑﻴﻤﺎﺭﻳﻬـﺎﯼ ﺗﻨﻔـﺴﯽ ﺍﺷـﺎﺭﻩ ﻧﻤـﻮﺩ‬
‫)ﺁﺭﺗــﻮﺭ ﻭ ﻫﻤﮑــﺎﺭﺍﻥ ‪ .(۱۹۸۸‬ﻋﻠــﯽ ﺭﻏــﻢ ﺁﻧﭽــﻪ ﮔﻔﺘــﻪ ﺷــﺪ‬
‫ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﺍﺭﺗﺒﺎﻃﯽ ﺑﺎ ﺩﻭ ﺑﺮﺍﺑـﺮ ﺷـﺪﻥ ﻣﺎﻫﻴﭽـﻪ ﻫـﺎ‬
‫ﻧﺪﺍﺭﺩ‪ ،‬ﺍﻣـﺎ ﺑـﻪ ﻧـﺴﺒﺖ‪ ،‬ﻳـﻚ ﺍﻓـﺰﺍﻳﺶ ﺩﺭ ﺗﻌـﺪﺍﺩ ﻓﻴﺒـﺮ ﻫـﺎﯼ‬
‫‪٢‬‬
‫ﻣﺎﻫﻴﭽﻪ ﺍﯼ )ﻫﻴﭙﺮ ﭘﻼﺯﻱ( ﻭ ﻃﻮﻳﻞ ﺷـﺪﻥ ﻓﻴﺒـﺮ ﻫـﺎ )ﻫـﺎﻳﭙﺮ‬
‫ﺗﺮﻭﻓﻲ(‪ ٣‬ﺭﺍ ﺍﻳﺠﺎﺩ ﻣﻲ ﻛﻨﺪ‪ .‬ﺩﺭ ﮔﺎﻭﻫـﺎﯼ ﻣﺎﻫﻴﭽـﻪ ﻣـﻀﺎﻋﻒ‬
‫ﺗﻌــﺪﺍﺩ ﻓﻴﺒــﺮ ﻫــﺎﯼ ﻣﺎﻫﻴﭽــﻪ ﺍﯼ ﺩﺭ ﺯﻣــﺎﻥ ﺗﻮﻟــﺪ ﺩﻭ ﺑﺮﺍﺑــﺮ‬
‫ﮔﺎﻭﻫﺎﯼ ﻋـﺎﺩﯼ ﺍﺳـﺖ‪ .‬ﮔﺎﻭﻫـﺎﯼ ﻣﺎﻫﻴﭽـﻪ ﻣـﻀﺎﻋﻒ ﺩﺍﺭﺍﯼ‬
‫ﺩﺭﺻﺪ ﺑﻴﺸﺘﺮﯼ ﺍﺯ ﻓﻴﺒﺮ ﻫﺎﯼ ﻣﺎﻫﻴﭽﻪ ﺍﯼ ﺳـﻔﻴﺪ ﻫـﺴﺘﻨﺪ‪ ،‬ﻭ‬
‫ﻣﻘﺪﺍﺭ ﻛﻼﮊﻥ ﻛﻤﺘـﺮﯼ ﺩﺍﺭﻧـﺪ‪ .‬ﻫﻤﭽﻨـﻴﻦ ﮔـﺰﺍﺭﺵ ﺷـﺪﻩ ﻛـﻪ‬
‫ﮔﺎﻭﻫــﺎﯼ ﺑــﺎ ﻣﺎﻫﻴﭽــﻪ ﻣــﻀﺎﻋﻒ ﺩﺍﺭﺍﯼ ﺑﺎﻓــﺖ ﻫــﺎﯼ ﻫﻤﺒﻨــﺪ‬
‫ﻛﻤﺘﺮﯼ ﻫﺴﺘﻨﺪ ﻛﻪ ﻣﯽ ﺗﻮﺍﻧﺪ ﺑﻪ ﺍﻓﺰﺍﻳﺶ ﺗﺮﺩﯼ ﮔﻮﺷﺖ ﻛﻤﻚ‬
‫ﻛﻨﺪ‪ .‬ﺍﻳﻦ ﻧﻜﺘﻪ ﺭﺍ ﺑﺎﻳﺪ ﻣﺘﺬﻛﺮ ﺷﺪ ﻛﻪ ﻋﻼﻭﻩ ﺑﺮ ﻧﮋﺍﺩ‪ ،‬ﺗﻐﺬﻳﻪ ﻭ‬
‫ﺟﻨﺲ ﻧﻴﺰ ﻓﻨﻮﺗﻴﭗ ﻣﺎﻫﻴﭽـﻪ ﻣـﻀﺎﻋﻒ ﺭﺍ ﺗﺤـﺖ ﺗـﺎﺛﻴﺮ ﻗـﺮﺍﺭ‬
‫ﻣﯽ ﺩﻫﻨـﺪ )ﺟـﺮﺍﺭﺩ ﻭ ﻫﻤﮑـﺎﺭﺍﻥ ‪ .(۱۹۹۱‬ﺣﻴﻮﺍﻧـﺎﺕ ﻣﺎﻫﻴﭽـﻪ‬
‫ﻣﻀﺎﻋﻒ ﮔﻮﺷﺖ ﻇﺮﻳﻒ ﺗﺮ ﻭ ﺗـﺮﺩ ﺗـﺮﯼ ﺗﻮﻟﻴـﺪ ﻣـﯽ ﻛﻨﻨـﺪ‪.‬‬
‫ﻫﻤﭽﻨﻴﻦ ﮔﻮﺷﺖ ﺣﻴﻮﺍﻧﺎﺕ ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﺑﻪ ﻃﻮﺭ ﻣﻌﻨـﯽ‬
‫ﺩﺍﺭﯼ ﺣﺎﻭﯼ ﻣﻘﺪﺍﺭ ﭼﺮﺑﯽ ﻛﻤﺘﺮﯼ ﺍﺳﺖ‪ .‬ﺍﻳﻦ ﻭﻳﮋﮔﻲ ﻫـﺎ ﺑـﻪ‬
‫ﻃﻮﺭ ﺯﻳﺎﺩﯼ ﻣﻨﻄﺒﻖ ﺑﺎ ﺍﺳﺘﺎﻧﺪﺍﺭﺩﻫﺎﯼ ﺳﻼﻣﺘﯽ ﺍﺳﺖ )ﺳـﻤﺖ‬
‫ﻣﻮﺭﺩ ﺍﺳﺘﻔﺎﺩﻩ ﻗﺮﺍﺭ ﮔﺮﻓﺘﻪ ﺍﺳﺖ‪ ،‬ﺍﺯﺟﻤﻠﻪ‪ DM ،‬ﻳﺎ ‪ D ،N‬ﻳـﺎ‬
‫‪2‬‬
‫‪Hyperplasia‬‬
‫‪3‬‬
‫‪Hypertrophy‬‬
‫ﺍﺭﺯﻳﺎﺑﯽ ﭼﻨﺪﺷﻜﻠﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﮋﺍﺩ ﺳﻨﺠﺎﺑﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ‪...‬‬
‫ﻣﻘﺪﻣﻪ‬
‫ﺷﻨﺎﺧﺖ ﺟﻨﺒﻪﻫﺎﻱ ﮊﻧﺘﻴﮑﻲ ﻭ ﮊﻥﻫـﺎﻱ ﻋﻤـﺪﻩ ﺗﺄﺛﻴﺮﮔـﺬﺍﺭ ﺑـﺮ‬
‫ﺗﻮﻟﻴﺪ ﮔﻮﺷﺖ ﺍﺧﻴﺮﺍ" ﻣﻮﺭﺩ ﺗﻮﺟﻪ ﻣﺤﻘﻘﺎﻥ ﮊﻧﺘﻴﮏ ﻭ ﺍﺻـﻼﺡ‬
‫ﻧﮋﺍﺩ ﻗـﺮﺍﺭ ﮔﺮﻓﺘـﻪ ﺍﺳـﺖ‪ .‬ﻣﻄﺎﻟﻌـﺎﺕ ﺑـﺮﺍﻱ ﻳـﺎﻓﺘﻦ ﮊﻥ ﻫـﺎﻱ‬
‫ﺑﻮﺟﻮﺩ ﺁﻭﺭﻧﺪﻩ ﻋﻀﻠﻪ ﻣﻀﺎﻋﻒ ﺩﺭ ﮔﺎﻭﻫﺎﻱ ﮔﻮﺷﺘﻲ ﻣﻨﺠﺮ‬
‫ﺑﻪ ﮐﺸﻒ ﮊﻥ ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ ﺷـﺪ ﮐـﻪ ﺑـﻪ ﻋﻨـﻮﺍﻥ ﮊﻥ ﮐﺎﻧﺪﻳـﺪﺍ‬
‫ﺑﺮﺍﻱ ﺻﻔﺖ ﺗﻮﻟﻴﺪ ﮔﻮﺷﺖ ﺑﺮﺭﺳﻲ ﻭ ﻧﻘﺶ ﺁﻥ ﺩﺭ ﻧﮋﺍﺩ ﻫﺎﻱ‬
‫ﻣﺨﺘﻠﻒ ﺩﺍﻡ ﻭ ﻃﻴﻮﺭ ﻭ ﺣﺘـﻲ ﺍﻧـﺴﺎﻥ ﺷـﻨﺎﺧﺘﻪ ﺷـﺪﻩ ﺍﺳـﺖ‪.‬‬
‫ﻣﻄﺎﻟﻌﺎﺕ ﻣﻠﮑﻮﻟﻲ ﮊﻥ ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ ﻧـﺸﺎﻥ ﺩﺍﺩﻩ ﮐـﻪ ﺍﻳـﻦ ﮊﻥ‬
‫ﭼﻨﺪ ﺷﮑﻞ ﻣﻲ ﺑﺎﺷﺪ )ﻣﺴﻌﻮﺩﻱ ﻭ ﻫﻤﮑﺎﺭﺍﻥ ‪۱۳۸۴‬؛ ﺑﻠﻴﻨﮓ ﻭ‬
‫ﻫﻤﮑﺎﺭﺍﻥ ‪ ۲۰۰۵‬ﻭ ﮐﺎﺳﺎﺱ ﻭ ﻫﻤﮑﺎﺭﺍﻥ ‪ .(۲۰۰۰‬ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ‬
‫ﻣﻬﺎﺭﻛﻨﻨﺪﻩ ﺭﺷﺪ ﻣﺎﻫﻴﭽـﻪ ﻫـﺎﻱ ﺍﺳـﮑﻠﺘﻲ ﻣـﻲ ﺑﺎﺷـﺪ ﻭ ﺍﮔـﺮ‬
‫ﺟﻬـﺸﻲ ﺩﺭ ﮊﻥ ﮐــﺪ ﮐﻨﻨـﺪﻩ ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ ﺍﺗﻔـﺎﻕ ﺍﻓﺘــﺪ‪ ،‬ﺑﺎﻋــﺚ‬
‫ﺗﻐﻴﻴــﺮ ﻧﻘــﺶ ﻣﻬــﺎﺭﻱ ﺁﻥ ﻭ ﺍﻓــﺰﺍﻳﺶ ﻋــﻀﻠﻪ ﻣــﻲ ﺷــﻮﺩ‬
‫)ﻣــﺴﻌﻮﺩﻱ ﻭ ﻫﻤﮑــﺎﺭﺍﻥ ‪۱۳۸۴‬؛ ﻓﻬﺮﺍﻧﮑــﺮﻭﮒ ﻭ ﻫﻤﮑــﺎﺭﺍﻥ‬
‫‪ ۱۹۹۹‬ﻭ ﺟﺎﻧﺴﻮﻥ ﻭ ﻫﻤﮑﺎﺭﺍﻥ ‪ .(۲۰۰۵‬ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺗﻌﻴـﻴﻦ‬
‫ﻧﻘــﺸﻪ ﻓﻴﺰﻳﻜــﻲ ﻛﻠــﻮﻥ ﻫــﺎﻱ ﻳــﺎﻙ‪ ١‬ﻣــﺸﺨﺺ ﺷــﺪ ﻛــﻪ ﮊﻥ‬
‫ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﮔﺎﻭ ﻧﺰﺩﻳﻚ ﺳـﺎﻧﺘﺮﻭﻣﺮ ﻛﺮﻭﻣـﻮﺯﻭﻡ ﺷـﻤﺎﺭﻩ ‪۲‬‬
‫ﻗﺮﺍﺭ ﺩﺍﺭﺩ ﻭ ﺟﺎﻳﮕﺎﻩ ﺁﻥ ﺩﺭ ﮔﻮﻧﻪ ﻫﺎﯼ ﻣﻄﺎﻟﻌﻪ ﺷﺪﻩ ﺍﺯ ﺟﻤﻠﻪ‬
‫ﺧﻮﻙ‪ ،‬ﮔﺎﻭﻣﻴﺶ‪ ،‬ﻣﺮﻍ ﻭ ﻣﻮﺵ ﺧﺎﻧﮕﯽ ﻣﺸﺨﺺ ﺷﺪﻩ ﺍﺳـﺖ‬
‫)ﮐــﺎﺑﻮﻻﮎ ﻭ ﮔــﺰﺍ ‪ .(۲۰۰۲‬ﮊﻥ ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ ﮔــﺎﻭﻱ ﺩﺍﺭﺍﻱ ‪۳‬‬
‫ﺍﮔـــﺰﻭﻥ ﻭ ‪ ۲‬ﺍﻳﻨﺘـــﺮﻭﻥ ﻣـــﻲ ﺑﺎﺷـــﺪ‪ .‬ﻣﻴﻮﺳـــﺘﺎﺗﻴﻦ ﺑﺎﻋـــﺚ‬
‫ﻣﻴﺎﻧﺠﻴﮕﺮﻱ ﺑﻴﺎﻥ ﮊﻥ ﺩﺭ ﻛﻨﺘﺮﻝ ﺷﻜﻞ ﻓﻴﺒﺮﻱ ﻣﺎﻫﻴﭽﻪ ﺍﺳـﺖ‬
‫ﻭ ﺑﺎ ﺟﻠﻮﮔﻴﺮﻱ ﺍﺯ ﺗﻜﺜﻴﺮ ﻣﻴﻮﺑﻼﺳﺘﻬﺎ‪ ،‬ﻋﻤﻼﹲ ﺭﺷـﺪ ﻋـﻀﻼﻧﻲ‬
‫ﺭﺍ ﻣﺘﻮﻗﻒ ﻣﻲ ﻛﻨﺪ‪ .‬ﺍﻳﻦ ﻋﻤـﻞ ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ ﺑﻄـﻮﺭ ﻋﻤـﺪﻩ ﺑـﻪ‬
‫ﺭﺷﺪ ﻣﺎﻫﻴﭽﻪ ﻫﺎ ﭘﻴﺶ ﺍﺯ ﺗﻮﻟﺪ‪ ،‬ﻳﻌﻨﻲ ﺯﻣـﺎﻥ ﺗﻜﺜﻴـﺮ ﻭ ﺗﻤـﺎﻳﺰ‬
‫ﻣﻴﻮﺑﻼﺳﺘﻬﺎ ﺑﺮﻣﻲ ﮔﺮﺩﺩ ﻭ ﺍﻧﺘﻈﺎﺭ ﻣﻲ ﺭﻭﺩ ﭼﻨﺎﻧﭽﻪ ﺟﻬـﺸﻲ‬
‫ﺑﺘﻮﺍﻧــﺪ ﺑﺎﻋــﺚ ﻛــﺎﻫﺶ ‪ mRNA‬ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ ﮔــﺮﺩﺩ‪ ،‬ﺭﺷــﺪ‬
‫ﻋﻀﻼﻧﻲ ﺑﻴﺸﺘﺮﻱ ﺩﺭ ﺣﻴﻮﺍﻥ ﻣﺸﺎﻫﺪﻩ ﺷـﻮﺩ )ﻣـﺴﻌﻮﺩﻱ ﻭ‬
‫ﻫﻤﮑــﺎﺭﺍﻥ ‪ .(۱۳۸۴‬ﻋﻼﺋــﻢ ﻣﺨﺘﻠﻔــﯽ ﺑــﺮﺍﯼ ﺗــﺸﺨﻴﺺ ﺑــﻴﻦ‬
‫ﺣﻴﻮﺍﻧﺎﺕ ﺑﺎ ﻓﻨﻮﺗﻴﭗ ﻣﺎﻫﻴﭽﻪ ﻣـﻀﺎﻋﻒ ﻭ ﺣﻴﻮﺍﻧـﺎﺕ ﻃﺒﻴﻌـﯽ‬
‫‪1‬‬
‫‪Yak‬‬
 ‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‪ /‬ﺟﻠﺪ ‪ ١٩‬ﺷﻤﺎﺭﻩ ‪ /١‬ﺳﺎﻝ ‪١٣٨٨‬‬
‫ﻧﮋﺍﺩ ﺑﻌﺪ ﺍﺯ ﮔﻮﺳﻔﻨﺪ ﺑﻠﻮﭼﻲ ﺩﺍﺭﺍﻱ ﺑﻴﺸﺘﺮﻳﻦ ﺗﻌﺪﺍﺩ ﺩﺭ ﺑـﻴﻦ‬
‫ﮔﻮﺳﻔﻨﺪﺍﻥ ﺍﻳﺮﺍﻥ ﺍﺳﺖ ﻭ ﺩﺭ ﺍﮐﺜﺮ ﻧﻘـﺎﻁ ﺍﺳـﺘﺎﻥ ﮐﺮﻣﺎﻧـﺸﺎﻩ‬
‫ﭘﺮﺍﮐﻨﺪﻩ ﺍﺳﺖ‪ .‬ﮐﻞ ﺟﻤﻌﻴﺖ ﺍﻳﻦ ﻧﮋﺍﺩ ﺑﺮ ﺍﺳـﺎﺱ ﺳﺮﺷـﻤﺎﺭﻱ‬
‫ﺳــﺎﻝ ‪ ۱۳۸۶‬ﺣــﺪﻭﺩ ‪ ۶۰۰۰۰۰‬ﺭﺍﺱ ﻣــﻲ ﺑﺎﺷــﺪ )ﻣﻮﻻﺋﻴــﺎﻥ‪،‬‬
‫‪ .(١٣٧٨‬ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﺍﻫﻤﻴـﺖ ﺍﻳـﻦ ﻧـﮋﺍﺩ ﺩﺭ ﺗﻮﻟﻴـﺪ ﮔﻮﺷـﺖ ﻭ‬
‫ﭘﺸﻢ ﺍﺳﺘﺎﻥ ﻭ ﻋﺪﻡ ﺍﻧﺠﺎﻡ ﺗﺤﻘﻴﻘﺎﺕ ﻣﻮﻟﮑﻮﻟﯽ‪ ،‬ﺑﻪ ﻭﻳﮋﻩ ﺭﻭﯼ‬
‫ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﻭ ﻧﻘﺶ ﺍﻳﻦ ﮊﻥ ﺩﺭ ﺗﻮﻟﻴﺪ ﮔﻮﺷـﺖ‪ ،‬ﻫـﺪﻑ ﺍﺯ‬
‫ﺍﻧﺠــﺎﻡ ﺗﺤﻘﻴـــﻖ ﺣﺎﺿـــﺮ ﺷﻨﺎﺳـــﺎﻳﻲ ﮊﻧﻮﺗﻴـــﭗ ﻫـــﺎﻱ ﮊﻥ‬
‫ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﻭ ﻓﺮﺍﻭﺍﻧـﻲ ﺁﻟـﻞ ﻫـﺎﻱ ﺁﻥ ﺑـﺮﺍﯼ ﺍﻭﻟـﻴﻦ ﺑـﺎﺭ ﺩﺭ‬
‫ﮔﻮﺳـﻔﻨﺪ ﻧــﮋﺍﺩ ﺳــﻨﺠﺎﺑﻲ ﺑــﻪ ﻛﻤــﻚ ﺭﻭﺵ ‪ PCR-RFLP‬ﻭ‬
‫ﺗﻌﻴﻴﻦ ﻣﻴﺰﺍﻥ ﭼﻨﺪﺷـﮑﻠﯽ ﺩﺭ ﺟﻤﻌﻴـﺖ ﮔﻮﺳـﻔﻨﺪﺍﻥ ﺳـﻨﺠﺎﺑﯽ‬
‫ﺑﻮﺩ‪.‬‬
‫ﻭﺍﻗﻊ ﺑﻴﻦ ﻧـﺸﺎﻧﮕﺮﻫﺎﻱ ‪ BM81124‬ﻭ ‪ BULGE20‬ﺛﺎﺑـﺖ‬
‫ﻣﻮﺍﺩ ﻭ ﺭﻭﺵﻫﺎ‬
‫ﺟﻬـﺖ ﺍﻧﺠـﺎﻡ ﺗﺤﻘﻴـﻖ‪ ،‬ﻧﻤﻮﻧـﻪ ﻫـﺎﻱ ﺧـﻮﻥ ﺍﺯ ‪ ۱۵۰‬ﺭﺍﺱ‬
‫ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﻲ ﺩﺭ ﺳﻄﺢ ﺍﺳﺘﺎﻥ ﻛﺮﻣﺎﻧـﺸﺎﻩ ﺑﺪﺳـﺖ ﺁﻣـﺪ‪.‬‬
‫ﺍﺳــﺘﺨﺮﺍﺝ ‪ DNA‬ﭘــﺲ ﺍﺯ ﺗﻐﻴﻴــﺮ ﻭ ﺑﻬﻴﻨــﻪﺳــﺎﺯﻱ ﺭﻭﺵ‬
‫ﺍﺳﺘﺨﺮﺍﺝ ﻧﻤﻜـﻲ )ﻣﻴﻠـﺮ ﻭ ﻫﻤﮑـﺎﺭﺍﻥ ‪ (۱۹۹۸‬ﺑـﻪ ﺷـﺮﺡ ﺯﻳـﺮ‬
‫ﺍﻧﺠﺎﻡ ﺷﺪ‪ ٤ :‬ﺳﻲ ﺳﻲ ﺧﻮﻥ ﻛﺎﻣﻞ ﺩﺍﺧﻞ ﻟﻮﻟﻪﺍﻱ ﭘﻼﺳـﺘﻴﻜﻲ‬
‫ﻣﻮﺳﻮﻡ ﺑﻪ ﻓﺎﻟﻜﻮﻥ ﺭﻳﺨﺘﻪ ﺷﺪ‪ .‬ﺩﻭ ﺑﺮﺍﺑﺮ ﺣﺠﻢ ﺧـﻮﻥ‪ ،‬ﺑـﺎﻓﺮ‬
‫ﺟﺪﺍ ﻛﻨﻨـﺪﻩ ) ‪5mM MgCl2, 0.32M Sucrose, Triton‬‬
‫‪ (X-100 1%, 10mM Tris-HCl pH=7.5,‬ﺍﺿـﺎﻓﻪ‬
‫ﮔﺮﺩﻳﺪ ﻭ ﺳﭙﺲ ﻭﺭﺗﻜﺲ ﺷﺪ‪ .‬ﻣﺤﻠﻮﻝ ﻓﻮﻕ ﺑﻪ ﻣﺪﺕ ‪ ٥‬ﺩﻗﻴﻘـﻪ‬
‫ﺑــﺎ ﺳــﺮﻋﺖ ‪ ٧٠٠٠‬ﺩﻭﺭ ﺩﺭ ﺩﻗﻴﻘــﻪ ﺳــﺎﻧﺘﺮﻳﻔﻴﻮﮊ ﺷــﺪ‪ .‬ﻣــﺎﻳﻊ‬
‫ﺭﻭﻳﻲ ﺑﻪ ﺁﺭﺍﻣﻲ ﺟﺪﺍ ﻭ ﺭﺳﻮﺏ ﺣﺎﺻﻠﻪ ﻧﮕـﻪ ﺩﺍﺷـﺘﻪ ﺷـﺪ‪٣ .‬‬
‫ﻣﻴﻠﯽ ﻟﻴﺘﺮ ﺑﺎﻓﺮ ﺟﺪﺍ ﻛﻨﻨﺪﻩ ﺑﻪ ﺭﺳﻮﺏ ﺣﺎﺻﻠﻪ ﺍﺿﺎﻓﻪ ﻭ ﻋﻤـﻞ‬
‫ﺳﺎﻧﺘﺮﻳﻔﻴﻮﮊ ﻭ ﺟﺪﺍﺳﺎﺯﯼ ﺭﺳﻮﺏ ﺗﻜﺮﺍﺭ ﺷﺪ ﺗﺎ ﺯﻣﺎﻧﻴﻜﻪ ﻳـﻚ‬
‫ﺭﺳﻮﺏ ﻛﺎﻣﻼﹰ ﺳﻔﻴﺪ ﺣﺎﺻﻞ ﺷﺪ‪ ٣ .‬ﻣﻴﻠﯽ ﻟﻴﺘﺮ ﺑﺎﻓﺮ ﻟﻴﺰ ﻛﻨﻨﺪﻩ‬
‫‪(pH=8.2 - 400mM NaCl, 2mM Na2EDTA,‬‬
‫)‪ 10mM Tris-Hcl‬ﺑــﻪ ﺭﺳــﻮﺏ ﺍﺿــﺎﻓﻪ ﻭ ﺑــﻪ ﺧــﻮﺑﻲ‬
‫ﻭﺭﺗﻜﺲ ﺷـﺪ‪ .‬ﻣﻘـﺪﺍﺭ ‪ ٢٠٠‬ﻣﻴﻜﺮﻭﻟﻴﺘـﺮ ‪ SDS1‬ﺩﻩ ﺩﺭﺻـﺪ ﻭ‬
‫)ﺑــﺎ ﻏﻠﻈــﺖ ‪٥٠‬‬ ‫‪2‬‬
‫‪ ٣٥٠‬ﻣﻴﻜﺮﻭﻟﻴﺘــﺮ ﺁﻧــﺰﻳﻢ ﭘﺮﻭﺗﺌﻴﻨــﺎﺯ‪K-‬‬
‫‪1‬‬
‫ﮔﻮﺳﻔﻨﺪﺍﻥ ﺳﻨﮕﻴﻦ ﻭﺯﻥ ﺍﻳﺮﺍﻥ ﺍﺳﺖ )ﻣﻮﻻﺋﻴﺎﻥ ‪ .(١٣٧٨‬ﺍﻳـﻦ‬
‫)‪Sodium Dodecyl Sulfate (SDS‬‬
‫‪2‬‬
‫‪Proteinase-K‬‬
‫ﺻﻮﻓﯽ‪ ،‬ﻣﺤﻤﺪﺁﺑﺎﺩﯼ ﻭ ‪...‬‬ ‫‪٨٤‬‬
‫ﻭ ﻫﻤﮑﺎﺭﺍﻥ ‪ .(۲۰۰۰‬ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺑﻴـﺸﺘﺮ ﺩﺭ ﮔـﺎﻭ ﻣـﻮﺭﺩ‬
‫ﻣﻄﺎﻟﻌــﻪ ﻗــﺮﺍﺭ ﮔﺮﻓﺘــﻪ ﺍﺳــﺖ ﻭ ﺩﺭ ﮔﻮﺳــﻔﻨﺪ ﮐﻤﺘــﺮ ﻣــﻮﺭﺩ‬
‫ﺑﺮﺭﺳﯽ ﻗـﺮﺍﺭ ﮔﺮﻓﺘـﻪ ﺍﺳـﺖ‪ .‬ﺗﺤﻘﻴﻘـﻲ ﺗﻮﺳـﻂ ﺟﺎﻧـﺴﻮﻥ ﻭ‬
‫ﻫﻤﮑﺎﺭﺍﻥ )‪ (۲۰۰۵‬ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧـﮋﺍﺩ ﺗﮑـﺴﻞ ﺍﻧﺠـﺎﻡ ﺷـﺪ‪ .‬ﺩﺭ‬
‫ﺍﻳــﻦ ﺗﺤﻘﻴــﻖ ﻧﺎﺣﻴــﻪ ﮊﻥ ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ ﺑــﺮ ﺭﻭﻱ ﮐﺮﻭﻣــﻮﺯﻡ‬
‫ﺷــﻤﺎﺭﻩ ‪ ۲‬ﮔﻮﺳــﻔﻨﺪ ﺟﻬــﺖ ﺷﻨﺎﺳــﺎﻳﻲ ﮊﻥ ﮐﺎﻧﺪﻳــﺪﺍ ﺑــﺮﺍﻱ‬
‫ﺻﻔﺎﺕ ﻻﺷﻪ ﻣﻮﺭﺩ ﺑﺮﺭﺳﻲ ﻗﺮﺍﺭ ﮔﺮﻓﺖ‪ .‬ﺩﺭ ﺍﻳﻦ ﭘﮋﻭﻫﺶ ﺍﺯ‬
‫‪ ۸‬ﻧﺸﺎﻧﮕﺮ ﻣﻴﮑﺮﻭﺳﺎﺗﻼﻳﺖ ﺟﻬـﺖ ﺗﻌﻴـﻴﻦ ﮊﻧﻮﺗﻴـﭗ ﺍﺳـﺘﻔﺎﺩﻩ‬
‫ﺷﺪﻩ ﻭ ﺩﺭ ﻧﻬﺎﻳﺖ ﺁﻧﺎﻟﻴﺰ ﺁﻣﺎﺭﻱ ﺑـﺮ ﺭﻭﻱ ﺩﺍﺩﻩ ﻫـﺎ ﺻـﻮﺭﺕ‬
‫ﮔﺮﻓـﺖ‪ .‬ﻧــﺸﺎﻧﻪ ﺍﻱ ﺍﺯ ﮊﻥ ﮐﺎﻧﺪﻳــﺪﺍ ﺑـﺮﺍﻱ ﺳــﺮﻋﺖ ﺭﺷــﺪ ﻳــﺎ‬
‫ﺻﻔﺎﺕ ﻻﺷﻪ ﺑﻪ ﺩﺳﺖ ﻧﻴﺎﻣﺪ‪ .‬ﺍﻣﺎ‪ ،‬ﺩﺭ ﻣﻮﺭﺩ ﺻﻔﺖ ﭼﺮﺑـﻲ ﻭ‬
‫ﻣﺎﻫﻴﭽﻪ ﺭﺍﻥ ﻭﺟﻮﺩ ﮊﻥ ﮐﺎﻧﺪﻳـﺪﺍ ﺑـﺮﺍﻱ ﺍﻓـﺮﺍﺩ ﻫﺘﺮﻭﺯﻳﮕـﻮﺕ‬
‫ﺷﺪ ﻭﻟﻲ ﺩﺭ ﻣﻮﺭﺩ ﺍﻓﺮﺍﺩ ﻫﻤﻮﺯﻳﮕﻮﺕ ﭼﻨﻴﻦ ﭼﻴﺰﻱ ﻣـﺸﺎﻫﺪﻩ‬
‫ﻧﺸﺪ‪ .‬ﻣـﺴﻌﻮﺩﻱ ﻭ ﻫﻤﮑـﺎﺭﺍﻥ )‪ (۱۳۸۴‬ﺑـﺮ ﺭﻭﻱ ﮔﻮﺳـﻔﻨﺪﺍﻥ‬
‫ﺑﻠﻮﭼﻲ ﺍﻳﺴﺘﮕﺎﻩ ﻋﺒﺎﺱ ﺁﺑـﺎﺩ ﻣـﺸﻬﺪ ﺑـﺎ ﺍﺳـﺘﻔﺎﺩﻩ ﺍﺯ ﺗﮑﻨﻴـﮏ‬
‫‪ PCR-SSCP‬ﭼﻨﺪﺷـــﮑﻠﯽ ﮊﻥ ﻣﻴﻮﺳـــﺘﺎﺗﻴﻦ ﺭﺍ ﺑﺮﺭﺳـــﻲ‬
‫ﮐﺮﺩﻧﺪ‪ ۱۱ .‬ﺟﻔﺖ ﺁﻏﺎﺯﮔﺮ ﺟﻬﺖ ﺍﻧﺠﺎﻡ ﻭﺍﮐﻨـﺸﻬﺎﻱ ‪ PCR‬ﺑـﺎ‬
‫ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺗﻮﺍﻟﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﮔـﺎﻭ ﻭ ﺑﺨـﺸﻲ ﺍﺯ ﺁﻥ ﺩﺭ‬
‫ﮔﻮﺳﻔﻨﺪ ﻃﺮﺍﺣﻲ ﺷﺪ‪ .‬ﺍﻳﻦ ‪ ۱۱‬ﺁﻏﺎﺯﮔﺮ ﺗﻘﺮﻳﺒـﺎ ﺗﻤـﺎﻣﻲ ﻃـﻮﻝ‬
‫ﮊﻥ ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ ﮔﻮﺳــﻔﻨﺪ ﺭﺍ ﭘﻮﺷــﺶ ﻣــﻲ ﺩﺍﺩﻧــﺪ‪ .‬ﻧﺘــﺎﻳﺞ‬
‫ﺣﺎﺻــﻞ ﺍﺯ ﺗﮑﺜﻴــﺮ ‪ ۶‬ﺁﻏــﺎﺯﮔﺮ ﻧــﺸﺎﻥ ﺩﺍﺩ ﮐــﻪ ﺗــﻮﺍﻟﻲ ﮊﻥ‬
‫ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﺍﻳﻦ ﺟﻤﻌﻴﺖ ﺑﺴﻴﺎﺭ ﺣﻔﺎﻇﺖ ﺷﺪﻩ ﻣﻲ ﺑﺎﺷـﺪ‪،‬‬
‫ﺩﺭ ﻣﻮﺭﺩ ‪ ۲‬ﺁﻏﺎﺯﮔﺮ ﺩﻳﮕﺮ ﻭ ﺩﻗﺖ ﺩﺭ ﺍﻟﮕﻮﻱ ﺑﺎﻧﺪﻱ ﺁﻧﻬـﺎ ﺑـﻪ‬
‫ﺍﻳﻦ ﻧﮑﺘﻪ ﺍﺷﺎﺭﻩ ﺷﺪ ﮐﻪ ﺟﻬﺸﻲ ﺗﮏ ﻧﻮﮐﻠﺌﻮﺗﻴﺪﻱ ﻭ ﻳﺎ ﺣـﺬﻑ‬
‫ﻭ ﺍﺿﺎﻓﻪ ﺩﺭ ﻳﮏ ﻣﺤﺪﻭﺩﻩ ﻣﻌﻴﻦ ﮊﻥ ﺍﺗﻔﺎﻕ ﺍﻓﺘـﺎﺩﻩ ﺍﺳـﺖ‪ .‬ﺩﺭ‬
‫ﺍﻳﻦ ﭘﮋﻭﻫﺶ ﺣـﺪﺍﻗﻞ ﻣﺮﺑﻌـﺎﺕ ﺍﺭﺯﺵ ﺍﺻـﻼﺣﻲ ﻭﺯﻥ ﺗﻮﻟـﺪ‬
‫ﺗﻔﺎﻭﺕ ﻣﻌﻨﻲ ﺩﺍﺭﻱ ﺍﺯ ﻟﺤﺎﻅ ﺁﻣﺎﺭﻱ ﺑﺎ ﮊﻧﻮﺗﻴﭙﻬـﺎﻱ ﻣﺨﺘﻠـﻒ‬
‫ﺣﺎﺻﻞ ﺍﺯ ﺗﮑﺜﻴﺮ ﻳﮏ ﺁﻏﺎﺯﮔﺮ ﭼﻨﺪ ﺷﮑﻞ ﻧﺸﺎﻥ ﺩﺍﺩ‪ ،‬ﻭﻟـﻲ ﺑـﺎ‬
‫ﺩﻳﮕﺮ ﺻﻔﺎﺕ ﺍﺭﺗﺒﺎﻁ ﻣﻌﻨﻲ ﺩﺍﺭﻱ ﻧﺪﺍﺷﺖ‪.‬‬
‫ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﻲ‪ ،‬ﺩﻧﺒﻪ ﺩﺍﺭ ﺑﺎ ﺟﺜﻪ ﺍﻱ ﺑﺰﺭﮒ ﻭ ﺩﺳﺖ ﻭ‬
‫ﭘﺎﻳﻲ ﺑﻠﻨـﺪ ﻣـﻲ ﺑﺎﺷـﺪ‪ .‬ﺍﻳـﻦ ﻧـﮋﺍﺩ ﺍﺯ ﻧﻈـﺮ ﺗﻮﻟﻴـﺪ ﮔﻮﺷـﺖ ﺍﺯ‬
 ‫‪٨٥‬‬ ‫ﺍﺭﺯﻳﺎﺑﯽ ﭼﻨﺪﺷﻜﻠﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﮋﺍﺩ ﺳﻨﺠﺎﺑﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ‪...‬‬

‫ﺣﺠﻢ ﻧﻬﺎﻳﻲ ‪ ۲۵‬ﻣﻴﮑﺮﻭﻟﻴﺘﺮ ﻭ ﻳﻚ ﺟﻔﺖ ﭘﺮﺍﻳﻤﺮ ﺑﺎ ﺗﻮﺍﻟﻲ ﺯﻳـﺮ‬ ‫ﻣﻴﻜﺮﻭﮔﺮﻡ ﺑﺮ ﻣﻴﻠﻲ ﻟﻴﺘﺮ( ﺑﻪ ﻣﺤﻠﻮﻝ ﺣﺎﺻـﻠﻪ ﺍﺿـﺎﻓﻪ ﺷـﺪ ﻭ‬
‫ﺍﺳﺘﻔﺎﺩﻩ ﮔﺮﺩﻳﺪ )ﺗﻴﻤﻮﺗﻲ ﻭ ﻫﻤﻜﺎﺭﺍﻥ‪.(۱۹۹۷ ،‬‬ ‫ﺩﺭ ﻃﻮﻝ ﺷﺐ ﺩﺭ ﺩﻣﺎﻱ ‪ ٣٧‬ﺩﺭﺟﻪ ﺳـﺎﻧﺘﻴﮕﺮﺍﺩ ﺩﺭ ﺣﻤـﺎﻡ ﺁﺏ‬
‫'‪5'-CCGGAGAGACTTTGG GCTTGA-3‬‬ ‫ﮔﺮﻡ ﻗﺮﺍﺭ ﺩﺍﺩﻩ ﺷﺪ ﺗﺎ ﻫﻀﻢ ﻣﺤﺘﻮﻳـﺎﺕ ﻫـﺴﺘﺔ ﮔﻠﺒـﻮﻝﻫـﺎﻱ‬
‫'‪5'-TCATGAGCACCCACAGCGGTC-3‬‬
‫ﺳﻔﻴﺪ ﻛﺎﻣﻞ ﮔـﺮﺩﺩ‪ .‬ﻳـﻚ ﻣﻴﻠـﻲ ﻟﻴﺘـﺮ ﻣﺤﻠـﻮﻝ ‪ NaCl‬ﺍﺷـﺒﺎﻉ‬
‫ﻭﺍﻛﻨﺶ ﺯﻧﺠﻴﺮﻩ ﺍﻱ ﭘﻠﻴﻤﺮﺍﺯ ﺑﺎ ﺑﺮﻧﺎﻣﻪ ﺣﺮﺍﺭﺗـﻲ ﺯﻳـﺮ ﺩﺭ‬
‫)ﺣﺪﻭﺩ ‪ ٦‬ﻣﻮﻻﺭ( ﺑﻪ ﻣﺤﻠﻮﻝ ﺍﺿﺎﻓﻪ ﮔﺮﺩﻳﺪ ﻭ ﺳـﭙﺲ ﻣﺤﻠـﻮﻝ‬
‫ﺩﺳﺘﮕﺎﻩ ﺗﺮﻣﻮﺳﺎﻳﻜﻠﺮ ﺍﻧﺠﺎﻡ ﺷـﺪ‪ :‬ﻭﺍﺳﺮﺷـﺖ ﺳـﺎﺯﻱ ﺍﻭﻟﻴـﻪ‬
‫ﺑﻪ ﻣﺪﺕ ‪ ١٥‬ﺛﺎﻧﻴﻪ ﺑﻪ ﺷﺪﺕ ﺗﻜﺎﻥ ﺩﺍﺩﻩ ﺷﺪ‪ .‬ﻣﺤﻠﻮﻝ ﻣﺰﺑﻮﺭ ﺑﻪ‬
‫‪ DNA‬ﺑﻪ ﻣﺪﺕ ‪ ۱‬ﺩﻗﻴﻘﻪ ﻭ ‪ ۹۴‬ﺩﺭﺟﻪ ﺳـﺎﻧﺘﻴﮕﺮﺍﺩ‪ ،‬ﺍﻧﺠـﺎﻡ ‪۳۳‬‬
‫ﻣﺪﺕ ‪ ١٥‬ﺩﻗﻴﻘﻪ ﺑﺎ ﺳﺮﻋﺖ ‪ ٢٥٠٠‬ﺩﻭﺭ ﺩﺭ ﺩﻗﻴﻘﻪ ﺳﺎﻧﺘﺮﻳﻔﻴﻮﮊ‬
‫ﺳﻴﮑﻞ ﺑﺎ ﻭﺍﺳﺮﺷﺖ ﺳﺎﺯﻱ ‪ DNA‬ﻃﻲ‪ ۳۰‬ﺛﺎﻧﻴﻪ ﻭ ‪ ۹۴‬ﺩﺭﺟﻪ‬
‫ﺷﺪ‪ .‬ﻣﺎﻳﻊ ﺭﻭﻳﻲ ﺑﻪ ﻳﻚ ﻟﻮﻟﺔ ﭘﻼﺳﺘﻴﻜﻲ ﺍﺳﺘﺮﻳﻞ ﺩﻳﮕﺮ ﺍﻧﺘﻘـﺎﻝ‬
‫ﺳﺎﻧﺘﻴﮕﺮﺍﺩ‪ ،‬ﺍﺗﺼﺎﻝ ﺁﻏﺎﺯﮔﺮ ﺑـﻪ ‪ DNA‬ﻃـﻲ ‪ ۶۰‬ﺛﺎﻧﻴـﻪ ﻭ ‪۵۸‬‬
‫ﺩﺍﺩﻩ ﺷﺪ‪ .‬ﺩﺭ ﺍﻳﻦ ﻣﺮﺣﻠﻪ ﭘﺮﻭﺗﺌﻴﻦﻫﺎﻱ ﺍﺗﺼﺎﻟﻲ ﺑﻪ ﺍﺳـﻴﺪﻫﺎﻱ‬
‫ﺩﺭﺟﻪ ﺳﺎﻧﺘﻴﮕﺮﺍﺩ ﻭ ﺑﺴﻂ ﺁﻏﺎﺯﮔﺮ ﻃﻲ ‪ ۲‬ﺩﻗﻴﻘﻪ ﻭ ‪ ۷۲‬ﺩﺭﺟـﻪ‬
‫ﻧﻮﻛﻠﺌﻴﻚ ﻫﻤﺮﺍﻩ ﺑﺎ ﻧﻤﻚ ﺩﺭ ﺍﻧﺘﻬﺎﻱ ﻟﻮﻟﻪ ﺭﺳﻮﺏ ﺳﻔﻴﺪ ﺭﻧﮕـﻲ‬
‫ﺳﺎﻧﺘﻴﮕﺮﺍﺩ ﻭ ﺑﺴﻂ ﺍﻧﺘﻬﺎﻳﻲ ‪ ۲‬ﺩﻗﻴﻘﻪ ﻭ ‪ ۷۲‬ﺩﺭﺟﻪ ﺳـﺎﻧﺘﻴﮕﺮﺍﺩ‪.‬‬
‫ﺗﺸﻜﻴﻞ ﺩﺍﺩﻧﺪ‪ .‬ﻫﻢ ﺣﺠﻢ ﻣﺤﻠﻮﻝ‪ ،‬ﻛﻠﺮﻭﻓﺮﻡ ﺍﺿـﺎﻓﻪ ﺷـﺪ ﻭ ﺑـﺎ‬
‫ﺟﻬﺖ ﻣﺸﺎﻫﺪﻩ ﻣﺤﺼﻮﻻﺕ ﻭﺍﻛﻨﺶ ﺯﻧﺠﻴـﺮﻩ ﺍﻱ ﭘﻠﻴﻤـﺮﺍﺯ‪ ،‬ﺍﺯ‬
‫ﺗﻜﺎﻥ ﺩﺍﺩﻥ ﺑﺎ ﻣﺤﻠﻮﻝ ﻣﺨﻠﻮﻁ ﺷﺪ‪ .‬ﺳﭙﺲ ﺑﻪ ﻣﺪﺕ ‪ ١٠‬ﺩﻗﻴﻘـﻪ‬
‫ﮊﻝ ﺁﮔﺎﺭﺯ ‪ %۱‬ﻭ ﻭﻟﺘﺎﮊ ‪ ۸۰‬ﻭﻟﺖ ﺑﻪ ﻣﺪﺕ ‪ ۱‬ﺳـﺎﻋﺖ ﺍﺳـﺘﻔﺎﺩﻩ‬
‫ﺩﺭ ‪ ٣٠٠٠‬ﺩﻭﺭ ﺩﺭ ﺩﻗﻴﻘﻪ ﺳﺎﻧﺘﺮﻳﻔﻮﮊ ﮔﺮﺩﻳـﺪ‪ .‬ﺁﻟـﻮﺩﮔﻲﻫـﺎ ﻭ‬
‫ﺷﺪ ﻭ ﺭﻧﮓ ﺁﻣﻴﺰﻱ ﮊﻝ ﺑﺎ ﺍﺗﻴـﺪﻳﻮﻡ ﺑﺮﻭﻣﺎﻳـﺪ ﺍﻧﺠـﺎﻡ ﮔﺮﻓـﺖ‪.‬‬
‫ﭘﺮﻭﺗﺌﻴﻦ ﺍﺿﺎﻓﻲ ﺩﺭ ﺍﻳﻦ ﻣﺮﺣﻠﻪ ﺟﺪﺍ ﺷﺪﻧﺪ‪ .‬ﻓﺎﺯ ﺁﺑـﻲ ﻛـﻪ ﺩﺭ‬
‫ﺳﭙﺲ ﻣﺤـﺼﻮﻻﺕ ﻭﺍﻛـﻨﺶ ﺯﻧﺠﻴـﺮﻩ ﺍﻱ ﭘﻠﻴﻤـﺮﺍﺯ ﺑـﻪ ﻛﻤـﻚ‬
‫ﺑﺎﻻ ﺗﺸﻜﻴﻞ ﺷﺪ ﺣﺎﻭﻱ ‪ DNA‬ﺑﻮﺩ ﻛﻪ ﺑﺎ ﺍﺣﺘﻴﺎﻁ ﺟـﺪﺍ ﻭ ﺑـﻪ‬
‫ﺁﻧﺰﻳﻢ ﺑﺮﺷﻲ ‪ HaeΙΙΙ‬ﻣﻮﺭﺩ ﻫﻀﻢ ﺁﻧﺰﻳﻤﻲ ﻗﺮﺍﺭ ﮔﺮﻓﺘﻨﺪ‪.‬‬
‫ﻟﻮﻟﻪ ﭘﻼﺳﺘﻴﻜﻲ ﺟﺪﻳﺪﻱ ﺍﻧﺘﻘﺎﻝ ﺩﺍﺩﻩ ﺷـﺪ‪ .‬ﺑـﺮﺍﻱ ﻣﺘـﺮﺍﻛﻢ ﺗـﺮ‬
‫ﺑﺮﺍﻱ ﻣﺸﺎﻫﺪﻩ ﻗﻄﻌﺎﺕ ﻫﻀﻢ ﺷﺪﻩ ﺍﺯ ﮊﻝ ﺁﻛﺮﻳﻼﻣﻴـﺪ ‪%۸‬‬
‫ﻧﻤﻮﺩﻥ ﺭﺷـﺘﻪ ﻫـﺎﻱ ‪ ،DNA‬ﺑـﻪ ﺍﻧـﺪﺍﺯﺓ ‪ ٠/١‬ﺣﺠـﻢ ﻣﺤﻠـﻮﻝ‬
‫ﻭ ﻭﻟﺘــﺎﮊ ‪ ۲۰۰‬ﺑــﻪ ﻣــﺪﺕ ‪ ۲‬ﺳــﺎﻋﺖ ﺍﺳــﺘﻔﺎﺩﻩ ﺷــﺪ ﻭ ﺑﻌــﺪ ﺍﺯ‬
‫ﺍﺳﺘﺎﺕ ﺳﺪﻳﻢ ‪ ٣‬ﻣـﻮﻻﺭ )‪ (PH=٥/٢‬ﺍﺿـﺎﻓﻪ ﺷـﺪ‪ .‬ﺩﻭ ﺑﺮﺍﺑـﺮ‬
‫ﺭﻧﮓ ﺁﻣﻴﺰﻱ ﺑﺎ ﺍﺗﻴﺪﻳﻮﻡ ﺑﺮﻭﻣﺎﻳﺪ‪ ،‬ﮊﻝ ﺍﺳﮑﻦ ﺷـﺪ ﻭ ﺧﻮﺍﻧـﺪﻥ‬
‫ﺣﺠﻢ ﻣﺤﻠﻮﻝ ﺑﺪﺳﺖ ﺁﻣﺪﻩ‪ ،‬ﺍﺗﺎﻧﻮﻝ ﻣﻄﻠﻖ ﺍﺿﺎﻓﻪ ﺷﺪ ﻭ ﻟﻮﻟـﻪ‬
‫ﺁﻟﻞ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﻧﺮﻡ ﺍﻓـﺰﺍﺭ ‪ Uvidoc‬ﺻـﻮﺭﺕ ﮔﺮﻓـﺖ ﮐـﻪ‬
‫ﺑﻪ ﺁﻫﺴﺘﮕﻲ ﭼﻨﺪ ﺑﺎﺭ ﻭﺍﺭﻭﻧﻪ ﺷﺪ ﺗﺎ ﺭﺷﺘﻪﻫﺎﻱ ‪ DNA‬ﻇﺎﻫﺮ‬
‫ﻧﻤﻮﻧﻪ ﺍﻱ ﺍﺯ ﺁﻥ ﺩﺭ ﺷﮑﻞ ‪ ۳‬ﻧـﺸﺎﻥ ﺩﺍﺩﻩ ﺷـﺪﻩ ﺍﺳـﺖ‪ .‬ﺑـﺮﺍﻱ‬
‫ﺷﺪﻧﺪ‪ .‬ﻛﻼﻑ ‪ DNA‬ﺑﺎ ﻣﻴﻠﺔ ﺷﻴﺸﻪﺍﻱ ﻛﻪ ﺩﺍﺭﺍﻱ ﺑـﺎﺭ ﻣﺜﺒـﺖ‬
‫ﺑــﺮﺁﻭﺭﺩ ﻓﺮﺍﻭﺍﻧــﻲ ﺁﻟــﻞ ﻫــﺎ‪ ،‬ﻣﺤﺎﺳــﺒﻪ ﻫﺘﺮﻭﺯﻳﮕﻮﺳــﻴﺘﻲ ﻭ‬
‫ﻣﻲﺑﺎﺷﺪ ﺟﻤﻊﺁﻭﺭﻱ ﺷﺪ ﻭ ﺑﻪ ﻣﺪﺕ ﺣﺪﻭﺩ ‪ ٥‬ﺗﺎ ‪ ١٠‬ﺩﻗﻴﻘﻪ ﺩﺭ‬
‫ﺁﺯﻣــﻮﻥ ﻣﺮﺑــﻊ ﮐــﺎ )‪ (χ2‬ﺍﺯ ﻧــﺮﻡ ﺍﻓــﺰﺍﺭ ‪POP Gene3.2‬‬
‫ﻣﺠﺎﻭﺭﺕ ﻫﻮﺍ ﺧﺸﻚ ﺷﺪ‪ DNA .‬ﺣﺎﺻﻠﻪ ﺩﻭ ﺑﺎﺭ ﺑـﺎ ﺍﺗـﺎﻧﻮﻝ‬
‫ﺍﺳﺘﻔﺎﺩﻩ ﮔﺮﺩﻳﺪ‪.‬‬
‫‪ ٧٠‬ﺩﺭﺻــﺪ ﺷﺴﺘــﺸﻮ ﺩﺍﺩﻩ ﺷــﺪ ﻭ ﻧﻬﺎﻳﺘ ـﺎﹰ ﺩﺭ ‪ ١٠٠‬ﺗــﺎ ‪٢٠٠‬‬
‫ﻣﻴﻜﺮﻭﻟﻴﺘـﺮ ﺍﺯ ﺑـﺎﻓﺮ ‪10mM Tris HCl, 0.2mM ) TE‬‬
‫ﻧﺘﺎﻳﺞ ﻭ ﺑﺤﺚ‬
‫‪ (pH=7.5 - Na2EDTA‬ﺣﻞ ﮔﺮﺩﻳـﺪ‪ .‬ﺗﻴـﻮﺏ ﻫـﺎﻱ ﺣـﺎﻭﻱ‬
‫ﺍﺳــﺘﻔﺎﺩﻩ ﺍﺯ ﺭﻭﺵ ﺍﺳــﺘﺨﺮﺍﺝ ﻧﻤﻜــﻲ ﺟﻬــﺖ ﺍﺳــﺘﺨﺮﺍﺝ‬
‫‪ ،DNA‬ﺑﻪ ﻣﺪﺕ ‪ ٢‬ﺳﺎﻋﺖ ﺩﺭ ﺩﻣـﺎﻱ ‪ ٣٧‬ﺩﺭﺟـﻪ ﺳـﺎﻧﺘﻴﮕﺮﺍﺩ‬
‫‪ DNA‬ﺍﺯ ﻧﻤﻮﻧﻪ ﺧﻮﻥ ﺑﺮﺗﺮﻱ ﺧـﻮﺑﻲ ﺭﺍ ﺍﺯ ﻟﺤـﺎﻅ ﻛﻤﻴـﺖ ﻭ‬
‫ﻧﮕﻬﺪﺍﺭﻱ ﺷﺪﻧﺪ ﺗﺎ ‪ DNA‬ﺑﻪ ﺧـﻮﺑﻲ ﺣـﻞ ﺷـﻮﺩ‪ .‬ﺳـﭙﺲ ﺩﺭ‬
‫ﻛﻴﻔﻴﺖ ﻭ ﺻﺮﻑ ﺯﻣـﺎﻥ ﻻﺯﻡ ﺩﺭ ﺍﺳﺘﺤـﺼﺎﻝ ‪ DNA‬ﻧـﺸﺎﻥ‬
‫ﺩﻣﺎﻱ ‪ -٢٠‬ﺩﺭﺟﻪ ﺳﺎﻧﺘﻴﮕﺮﺍﺩ ﺫﺧﻴﺮﻩ ﺷﺪﻧﺪ‪ .‬ﺗﻌﻴﻴﻦ ﮐﻴﻔﻴـﺖ ﻭ‬
‫ﺩﺍﺩ‪ .‬ﺗﻜﺜﻴﺮ ﻗﻄﻌـﻪ ‪ ۳۳۷‬ﺟﻔـﺖ ﺑـﺎﺯﻱ ﺍﺯ ﮊﻥ ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ ﺑـﻪ‬
‫ﮐﻤﻴﺖ ‪ DNA‬ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺭﻭﺵ ﺍﻟﻜﺘﺮﻭﻓـﻮﺭﺯ ﻣﻘﺎﻳـﺴﻪ ﺍﻱ‬
‫ﻛﻤﻚ ﻭﺍﻛﻨﺶ ﺯﻧﺠﻴﺮﻩ ﺍﻱ ﭘﻠﻴﻤﺮﺍﺯ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺁﻏﺎﺯﮔﺮﻫﺎﻱ‬
‫ﺑﺮ ﺭﻭﻱ ﮊﻝ ﺁﮔﺎﺭﺯ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﻣﻘﺎﺩﻳﺮ ﻣﺸﺨـﺼﻲ ‪DNA‬‬
‫ﺍﺧﺘﺼﺎﺻﻲ ﺑﻪ ﺧﻮﺑﻲ ﺻﻮﺭﺕ ﮔﺮﻓﺖ ﻭ ﺩﺭ ﻧﺘﻴﺠـﻪ ﺍﺳـﺘﻔﺎﺩﻩ‬
‫ﻓـﺎﮊ ﻻﻣﺒـﺪﺍ ﺍﻧﺠــﺎﻡ ﺷـﺪ‪ .‬ﺟﻬـﺖ ﺍﻧﺠــﺎﻡ ﻭﺍﻛـﻨﺶ ﺯﻧﺠﻴــﺮﻩ ﺍﻱ‬
‫ﺍﺯ ﺑﺮﻧﺎﻣــﻪ ﺣﺮﺍﺭﺗــﻲ ﻣﻨﺎﺳــﺐ‪ ،‬ﺁﻏﺎﺯﮔﺮﻫــﺎﻱ ﺍﺧﺘــﺼﺎﺻﻲ ﻭ‬
‫ﭘﻠﻴﻤﺮﺍﺯ ﺍﺯ ﻛﻴﺖ ‪) PCR Master kit‬ﺷـﺮﮐﺖ ﺳـﻴﻨﺎ ﮊﻥ( ﺩﺭ‬
‫ﺷﺮﺍﻳﻂ ﺧﻮﺏ ﺁﺯﻣﺎﻳﺸﮕﺎﻫﻲ ﻛﻪ ﻓﺮﺍﻫﻢ ﺷﺪ ﻗﻄﻌﻪ ‪ ۳۳۷‬ﺟﻔـﺖ‬
 ‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‪ /‬ﺟﻠﺪ ‪ ١٩‬ﺷﻤﺎﺭﻩ ‪ /١‬ﺳﺎﻝ ‪١٣٨٨‬‬
‫ﻧﺸﺎﻥ ﺩﻫﻨﺪﻩ ﺣﻀﻮﺭ ﺑﻌﻀﻲ ﻋﻮﺍﻣﻞ ﺑﺮ ﻫﻢ ﺯﻧﻨﺪﻩ ﺗﻌﺎﺩﻝ‪ ،‬ﺍﺯ‬
‫ﺟﻤﻠﻪ ﻣﻬﺎﺟﺮﺕ ﻭ ﺍﻧﺘﺨﺎﺏ ﺍﺳﺖ‪ ،‬ﮐﻪ ﻣﻬﺎﺟﺮﺕ ﺧـﺼﻮﺻﺎﹰ ﺩﺭ‬
‫ﻣـﻮﺭﺩ ﻧﺮﻫـﺎﻳﻲ ﻛـﻪ ﺍﺯ ﺧـﺎﺭﺝ ﮔﻠـﻪ ﻭﺍﺭﺩ ﻣـﻲ ﺷـﻮﻧﺪ ﻭ ﻳـﮏ‬
‫ﺟﺮﻳﺎﻥ ﮊﻧﻲ ﺍﻳﺠﺎﺩ ﻣﯽ ﮐﻨﻨـﺪ ﺻـﺪﻕ ﻣـﻲ ﮐﻨـﺪ‪ .‬ﺍﻟﺒﺘـﻪ ﺍﻧـﺪﺍﺯﻩ‬
‫ﻧﻤﻮﻧﻪ ﻣﻮﺭﺩ ﺑﺮﺭﺳﯽ ﻫﻢ ﺩﺭ ﺍﻳﻦ ﺍﻣﺮ ﻣﯽ ﺗﻮﺍﻧﺪ ﻣﻮﺛﺮ ﺑﺎﺷﺪ‪.‬‬
‫ﻣﻴﺰﺍﻥ ﻫﺘﺮﻭﺯﻳﮕﻮﺳﻴﺘﻲ ﺑﺮﺁﻭﺭﺩ ﺷﺪﻩ ﺑﻮﺳﻴﻠﻪ ﺷـﺎﺧﺺ‬
‫‪٣‬‬
‫ﻧﺌﯽ ﺩﺭ ﺟﻤﻌﻴﺖ ‪ ۰/۰۵۱۹‬ﺑﺮﺁﻭﺭﺩ ﮔﺮﺩﻳﺪ‪ .‬ﻛـﻪ ﺍﻳـﻦ ﭘـﺎﺭﺍﻣﺘﺮ‬
‫ﺣــﺎﻛﻲ ﺍﺯ ﺗﻨــﻮﻉ ﮊﻧﺘﻴﻜــﻲ ﭘــﺎﻳﻴﻦ ﺍﻳــﻦ ﺟﻤﻌﻴــﺖ ﺍﺯ ﻧﻈــﺮ ﮊﻥ‬
‫ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺍﺳﺖ‪ .‬ﻣﻨﺎﺑﻊ ﻣﺨﺘﻠﻒ‪ ،‬ﺁﻟﻞ ‪ M‬ﺭﺍ ﺑـﻪ ﻋﻨـﻮﺍﻥ ﺁﻟـﻞ‬
‫ﻣﻄﻠﻮﺏ ﺟﻬﺖ ﺍﺻﻼﺡ ﻧﮋﺍﺩ ﻭ ﺑﻬﺒﻮﺩ ﻛﻴﻔﻴﺖ ﻭ ﻛﻤﻴﺖ ﮔﻮﺷـﺖ‬
‫ﻣﻌﺮﻓﻲ ﻛﺮﺩﻩ ﺍﻧﺪ‪ .‬ﺩﺭﺳـﺖ ﺍﺳـﺖ ﮐـﻪ ﻓﺮﺍﻭﺍﻧـﻲ ﺍﻳـﻦ ﺁﻟـﻞ ﺩﺭ‬
‫ﺟﻤﻌﻴﺖ ﺑﺴﻴﺎﺭ ﭘﺎﻳﻴﻦ ﺍﺳﺖ‪ ،‬ﻭﻟﯽ ﺑﺎﻳﺪ ﺗﻮﺟـﻪ ﺩﺍﺷـﺖ ﮐـﻪ ﻣـﺎ‬
‫ﻓﻘﻂ ﻳﮏ ﺟﻬﺶ ﮐﻪ ﺑﺎﻋـﺚ ﻣﺎﻫﻴﭽـﻪ ﻣـﻀﺎﻋﻒ ﻣـﯽ ﺷـﻮﺩ ﺭﺍ‬
‫ﺑﺮﺭﺳﯽ ﮐﺮﺩﻳﻢ‪ ،‬ﺩﺭ ﺣﺎﻟﯽ ﮐﻪ ﺟﻬـﺶ ﻫـﺎﯼ ﻣﺘﻔـﺎﻭﺗﯽ ﺑﺎﻋـﺚ‬
‫ﻣﺎﻫﻴﭽﻪ ﻣـﻀﺎﻋﻒ ﻣـﯽ ﺷـﻮﻧﺪ‪ .‬ﺑـﺮﺍﯼ ﻧﻤﻮﻧـﻪ‪ ،‬ﺍﻟﮑـﺴﺎﻧﺪﺭ ﻭ‬
‫ﻫﻤﮑﺎﺭﺍﻥ )‪ (١٩٩٧‬ﺑﺎ ﻣﻄﺎﻟﻌﻪ ﻣﻠﮑﻮﻟﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﻧـﺸﺎﻥ‬
‫ﺩﺍﺩﻧــﺪ ﮐــﻪ ﻳــﮏ ﺟﻬــﺶ ﺩﺭ ﮔــﺎﻭ ﻣــﺴﺌﻮﻝ ﺻــﻔﺖ ﻣﺎﻫﻴﭽــﻪ‬
‫ﻣﻀﺎﻋﻒ ﻣﻲ ﺑﺎﺷﺪ‪ .‬ﺁﻧﻬﺎ ﺑﺎ ﻣﻘﺎﻳﺴﻪ ﻣﻠﮑﻮﻟﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ‬
‫ﺩﺭ ﮔـﺎﻭ ﻧــﮋﺍﺩ ﻫﻠــﺸﺘﺎﻳﻦ ﻭ ﭘﻴــﺪﻣﺎﻧﺘﺲ‪ ،‬ﮐــﻪ ﻓﺮﺍﻭﺍﻧــﻲ ﻭﻗــﻮﻉ‬
‫ﻣﺎﻫﻴﭽﻪ ﻣـﻀﺎﻋﻒ ﺩﺭ ﺁﻥ ﺯﻳـﺎﺩ ﺍﺳـﺖ ﺩﺭﻳﺎﻓﺘﻨـﺪ ﮐـﻪ ﺩﺭ ﺩﻭ‬
‫ﻧﻮﮐﻠﺌﻮﺗﻴﺪ ﺑﺎ ﻫﻢ ﺍﺧﺘﻼﻑ ﺩﺍﺭﻧﺪ‪ .‬ﺩﺭ ﺍﮔـﺰﻭﻥ ﺷـﻤﺎﺭﻩ ﻳـﮏ ‪C‬‬
‫ﺑﻪ ‪ A‬ﻭ ﺑﺎﻟﻄﺒﻊ ﻟﻮﺳﻴﻦ ﺑﻪ ﻓﻨﻴﻞ ﺁﻻﻧﻴﻦ ﻭ ﺩﺭ ﺍﮔﺰﻭﻥ ﺷﻤﺎﺭﻩ‬
‫‪ ٣‬ﺟﻬﺶ ‪ G‬ﺑﻪ ‪ A‬ﻭ ﺑـﺎﻟﻄﺒﻊ ﺳﻴـﺴﻴﺘﻦ ﺑـﻪ ﺗﻴـﺮﻭﺯﻳﻦ ﺗﺒـﺪﻳﻞ‬
‫ﻣﻲ ﺷﻮﺩ ﻭ ﺁﺭﺗﻮﺭ ﻭ ﻫﻤﮑﺎﺭﺍﻥ )‪ (١٩٨٨‬ﻧﻴﺰ ﻧﺸﺎﻥ ﺩﺍﺩﻧﺪ ﮐـﻪ‬
‫ﺣﺬﻑ ‪ ١١‬ﻧﻮﮐﻠﺌﻮﺗﻴﺪ ﺩﺭ ﺍﮔﺰﻭﻥ ﺷـﻤﺎﺭﻩ ‪ ٣‬ﻧـﮋﺍﺩ ﺑﻠـﮋﻳﻦ ﺑﻠـﻮ‬
‫ﺭﺥ ﺩﺍﺩﻩ ﺍﺳﺖ‪ .‬ﺑﻨﺎﺑﺮﺍﻳﻦ‪ ،‬ﺑﺮﺍﯼ ﺍﻃﻤﻴﻨﺎﻥ ﺍﺯ ﻧﺘﻴﺠﻪ ﮔﻴﺮﯼ ﺑﺎﻳـﺪ‬
‫ﺗﻤﺎﻣﯽ ﺟﻬﺶ ﻫﺎ ﺭﺍ ﺑﺮﺭﺳﯽ ﻧﻤﻮﺩ‪.‬‬
‫ﻧﻈﺮ ﺑﻪ ﺍﻳﻦ ﮐﻪ ﺻﻔﺎﺕ ﻛﻤﻲ ﺗﻮﺳﻂ ﺑﺮﺁﻳﻨﺪ ﺗﻌﺪﺍﺩ ﺯﻳـﺎﺩﻱ‬
‫ﮊﻥ ﺑﺎ ﺍﺛﺮﺍﺕ ﮐﻢ ﻭ ﻫﻤﭽﻨﻴﻦ ﺍﺛﺮ ﻣﺘﻘﺎﺑﻞ ﺑﻴﻦ ﺁﻧﻬﺎ ﻛﻨﺘـﺮﻝ ﻣـﻲ‬
‫ﺷﻮﺩ‪ ،‬ﻟﺬﺍ ﻧﺎﻣﻨﺎﺳﺐ ﺑﻮﺩﻥ ﻓﺮﺍﻭﺍﻧـﻲ ﺁﻟـﻞ ﻣﻄﻠـﻮﺏ ﺩﺭ ﺳـﻄﺢ‬
‫ﻳﻚ ﻟﻮﻛﻮﺱ ﻧﻤﻲ ﺗﻮﺍﻧﺪ ﮔـﻮﺍﻫﻲ ﺑـﺮ ﻋﻤﻠﻜـﺮﺩ ﻧـﺎﻣﻄﻠﻮﺏ ﻳـﻚ‬
‫‪1‬‬
‫‪3‬‬
‫‪Nei‬‬
‫ﺻﻮﻓﯽ‪ ،‬ﻣﺤﻤﺪﺁﺑﺎﺩﯼ ﻭ ‪...‬‬ ‫‪٨٦‬‬
‫ﺑﺎﺯﻱ ﺑﺪﻭﻥ ﻗﻄﻌﺎﺕ ﻏﻴﺮ ﺍﺧﺘﺼﺎﺻﻲ ﺑﺪﺳﺖ ﺁﻣـﺪ )ﺷـﮑﻞ ‪(۲‬‬
‫ﻛﻪ ﺑﺎ ﻧﺘﺎﻳﺞ ﺗﻴﻤـﻮﺗﻲ ﻭ ﻫﻤﻜـﺎﺭﺍﻥ )‪ (۱۹۹۷‬ﺑـﺮ ﺭﻭﻱ ﺍﻧـﺴﺎﻥ‪،‬‬
‫ﮔﺎﻭ‪ ،‬ﮔﻮﺳﻔﻨﺪ ﻭ ﺑﺰ ﻣﻄﺎﺑﻘﺖ ﺩﺍﺷﺖ‪.‬‬
‫ﮔﻮﺳــﻔﻨﺪﺍﻥ ﺑــﺎ ﮊﻧﻮﺗﻴــﭗ ﻫﺘﺮﻭﺯﻳﮕــﻮﺕ )‪ (Mm‬ﺩﺍﺭﺍﻱ‬
‫ﻗﻄﻌﺎﺕ ‪ ۱۳۱ ، ۱۲۳، ۸۳‬ﻭ ﻗﻄﻌﻪ ﺑﺮﺵ ﻧﺨـﻮﺭﺩﻩ ‪ ۳۳۷‬ﺟﻔـﺖ‬
‫ﺑــﺎﺯﻱ ﺑﻮﺩﻧــﺪ‪ .‬ﺩﺭ ﻫﻤﻮﺯﻳﮕــﻮﺕ ﻏﺎﻟــﺐ )‪ (MM‬ﻗﻄﻌــﻪ ‪۳۳۷‬‬
‫ﺟﻔﺖ ﺑﺎﺯﻱ ﺑﺪﻭﻥ ﺑﺮﺵ ﺑﺎﻗﻲ ﻣﺎﻧﺪ‪ .‬ﺩﺭ ﻫﻤﻮﺯﻳﮕـﻮﺕ ﻣﻐﻠـﻮﺏ‬
‫)‪ (mm‬ﻧﻴﺰ ﻗﻄﻌـﺎﺕ ‪ ۱۲۳، ۸۳‬ﻭ‪ ۱۳۱‬ﺟﻔـﺖ ﺑـﺎﺯﻱ ﻣـﺸﺎﻫﺪﻩ‬
‫ﺷـــﺪ‪ .‬ﺑﻌـــﺪ ﺍﺯ ﺑـــﺮﺁﻭﺭﺩ ﻓﺮﺍﻭﺍﻧـــﻲ ﺁﻟـــﻞ ﻫـــﺎ‪ ،‬ﻣﺤﺎﺳـــﺒﻪ‬
‫ﻫﺘﺮﻭﺯﻳﮕﻮﺳﻴﺘﻲ ﻭ ﺁﺯﻣﻮﻥ ﻣﺮﺑﻊ ﮐﺎ )‪ (χ2‬ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﻧﺮﻡ‬
‫ﺍﻓــﺰﺍﺭ ‪ PopGene‬ﻣــﺸﺨﺺ ﮔﺮﺩﻳــﺪ ﮐــﻪ ﺁﻟــﻞ ‪ m‬ﺩﺍﺭﺍﻱ‬
‫ﺑﻴﺸﺘﺮﻳﻦ ﻓﺮﺍﻭﺍﻧﻲ )‪ (۰/۹۷‬ﻭ ﺁﻟﻞ ‪ M‬ﺩﺍﺭﺍﻱ ﻛﻤﺘﺮﻳﻦ ﻓﺮﺍﻭﺍﻧﻲ‬
‫)‪(۰/۰۳‬ﺑﻮﺩﻧــﺪ )ﺟــﺪﻭﻝ ‪ .(۱‬ﺍﻳــﻦ ﻧﺘــﺎﻳﺞ ﺑــﺎ ﻧﺘــﺎﻳﺞ ﭘــﮋﻭﻫﺶ‬
‫ﺩﻭﺭﺍﻙ ﻭ ﻫﻤﻜـــﺎﺭﺍﻥ )‪ (۲۰۰۲‬ﺑـــﺮ ﺭﻭﻱ ﮔﺎﻭﻫـــﺎﻱ ﻧـــﮋﺍﺩ‬
‫ﻛﺎﺭﻭﻟﻴﺎﺱ‪ ١‬ﻣﻄﺎﺑﻘﺖ ﺩﺍﺷﺖ ﻭﻟﻲ ﺑﺎ ﻧﺘـﺎﻳﺞ ﻭﻟـﺮ ﻭ ﻫﻤﻜـﺎﺭﺍﻥ‬
‫)‪ (۲۰۰۱‬ﺩﺭ ﻧﮋﺍﺩﻫﺎﻱ ﮔﺎﻭ ﭘﻴﺪﻣﺎﻧﺘﺲ‪ ٢‬ﻣﻐﺎﻳﺮﺕ ﺩﺍﺷﺖ‪ .‬ﺩﻟﻴـﻞ‬
‫ﺍﻳـﻦ ﺍﻣــﺮ ﺑـﻪ ﻭﻗــﻮﻉ ﺑــﺎﻻﻱ ﻓﻨﻮﺗﻴـﭗ ﻣﺎﻫﻴﭽــﻪ ﻣــﻀﺎﻋﻒ ﺩﺭ‬
‫ﮔﺎﻭﻫﺎﻱ ﭘﻴﺪﻣﺎﻧﺘﺲ ﻣﺮﺑﻮﻁ ﻣﻲﺷﻮﺩ‪ .‬ﻫﻤﭽﻨﻴﻦ ﻧﺘﺎﻳﺞ ﺣﺎﺻﻞ‬
‫ﺍﺯ ﺍﻳﻦ ﭘﮋﻭﻫﺶ ﺑﺎ ﻧﺘﺎﻳﺞ ﺑﻪ ﺩﺳﺖ ﺁﻣﺪﻩ ﺩﺭ ﻧﮋﺍﺩ ﮔـﺎﻭ ﺑﻠـﮋﻳﻦ‬
‫ﺑﻠﻮ‪ ،‬ﻛﻪ ﻓﻨﻮﺗﻴﭗ ﻣﺎﻫﻴﭽﻪ ﻣﻀﺎﻋﻒ ﺩﺭ ﺁﻥ ﺑﻪ ﻭﻓﻮﺭ ﻣـﺸﺎﻫﺪﻩ‬
‫ﻣــﻲ ﺷــﻮﺩ ﻣﻐــﺎﻳﺮﺕ ﺩﺍﺭﺩ )ﺍﻟﮑــﺴﺎﻧﺪﺭﺍ ﻭ ﻫﻤﮑــﺎﺭﺍﻥ‪۱۹۹۷ ،‬؛‬
‫ﮐﺎﺳﺎﺱ ﻭ ﻫﻤﮑـﺎﺭﺍﻥ‪ .(۲۰۰۰ ،‬ﺍﺯ ﺁﻥ ﺟـﺎﻳﻲ ﻛـﻪ ﻫـﻴﭻ ﮔﻮﻧـﻪ‬
‫ﺗﺤﻘﻴﻘﻲ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﺑـﻪ ﺷـﻜﻞ ﻓـﻮﻕ ﮔـﺰﺍﺭﺵ ﻧـﺸﺪﻩ ﺍﺳـﺖ‬
‫ﻧﻤﻲﺗﻮﺍﻥ ﻣﻘﺎﻳﺴﺎﺕ ﺟﺎﻟﺐ ﺗﻮﺟﻬﻲ ﺭﺍ ﺩﺭ ﺍﻳﻦ ﺧﺼﻮﺹ ﺑﻴﺎﻥ‬
‫ﻛﺮﺩ‪ .‬ﺍﺯ ﻃﺮﻓﯽ‪ ،‬ﻧﻤﯽ ﺗـﻮﺍﻥ ﮔﻔـﺖ ﻛـﻪ ﮊﻥ ﻣﻴﻮﺳـﺘﺎﺗﻴﻦ ﻭ ﻳـﺎ‬
‫ﺟﺎﻳﮕﺎﻫﻲ ﻧﺰﺩﻳﻚ ﺑﻪ ﺁﻥ ﺩﺭ ﻣﺎﻫﻴﭽﻪﺩﺍﺭ ﺷﺪﻥ ﮔﻮﺳﻔﻨﺪ ﻣﻮﺛﺮ‬
‫ﻧﻤﻲ ﺑﺎﺷﺪ‪ .‬ﺑﺮﺍﯼ ﺍﻃﻤﻴﻨﺎﻥ ﮐﺎﻣﻞ ﺑﺎﻳﺪ ﻣﻄﺎﻟﻌﺎﺕ ﺗﻜﻤﻴﻠﯽ ﺍﻧﺠﺎﻡ‬
‫ﺷﻮﺩ ﻭ ﺍﺭﺗﺒﺎﻁ ﺁﻥ ﺩﺭ ﺗﻌﺎﻣـﻞ ﺑـﺎ ﺟﺎﻳﮕـﺎﻩﻫـﺎﻱ ﺩﻳﮕـﺮ ﮊﻧـﻲ‬
‫ﺑﺮﺭﺳﯽ ﺷﻮﺩ‪ .‬ﺁﺯﻣﻮﻥ ﻣﺮﺑﻊ ﻛﺎ ﻧﺸﺎﻥ ﺩﺍﺩ ﻛﻪ ﺗﻌﺎﺩﻝ ﻫﺎﺭﺩﻱ‬
‫ﻭﻳﻨﺒــﺮﮒ ﺩﺭ ﺍﻳــﻦ ﺟﻤﻌﻴــﺖ ﺑــﺮﺍﻱ ﺟﺎﻳﮕــﺎﻩ ﮊﻥ ﻣﻴﻮﺳــﺘﺎﺗﻴﻦ‬
‫ﺑﺮﻗﺮﺍﺭ ﻧﻴﺴﺖ )‪ .(P<۰/۰۵‬ﻋﺪﻡ ﺗﻌﺎﺩﻝ ﺟﺎﻳﮕـﺎﻩ ﻫـﺎ ﺍﺣﺘﻤـﺎﻻ"‬
‫‪Charolias‬‬
‫‪2‬‬
‫‪Piedomontese‬‬
 ‫‪٨٧‬‬ ‫ﺍﺭﺯﻳﺎﺑﯽ ﭼﻨﺪﺷﻜﻠﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﮋﺍﺩ ﺳﻨﺠﺎﺑﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ‪...‬‬

‫ﺻﻔﺖ ﺩﺭ ﻳﻚ ﻧﮋﺍﺩ ﺑﺎﺷﺪ ﻭ ﺑﺎﻟﻄﺒﻊ ﺑﺮﺭﺳﻲ ﻭﺿﻌﻴﺖ ﺟﺎﻳﮕﺎﻩ‬

‫ﻫﺎﻱ ﮊﻧﻲ ﺩﻳﮕﺮ ﻧﻴﺰ ﻣﻮﺭﺩ ﻧﻴﺎﺯ ﺍﺳﺖ‪.‬‬
‫ﺩﺭ ﻛــﻞ ﺟﻤﻌﻴــﺖ‪ ،‬ﻣﻘــﺪﺍﺭ ﺷــﺎﺧﺺ ﺷــﺎﻧﻮﻥ )‪ (I‬ﺑﺮﺍﺑــﺮ‬
‫‪ ۰/۱۲۳‬ﻭ ﻣﻴﺎﻧﮕﻴﻦ ﻫﺘﺮﻭﺯﻳﮕﻮﺗﯽ ﭘﺎﻳﻴﻦ ﻭ ﺑﺮﺍﺑـﺮ ‪ ۰/۰۵‬ﺑـﻮﺩ‪،‬‬
‫ﮐﻪ ﻧﺸﺎﻥ ﺩﻫﻨﺪﻩ ﻭﺟﻮﺩ ﭼﻨﺪ ﺷﮑﻠﯽ ﺩﺭ ﺍﻳﻦ ﻧـﮋﺍﺩ ﺍﺳـﺖ‪ .‬ﺍﻳـﻦ‬
‫ﺍﻣﺮ ﻧﺸﺎﻥ ﺩﻫﻨﺪﻩ ﺍﻳـﻦ ﺍﺳـﺖ ﮐـﻪ ﻣـﯽ ﺗـﻮﺍﻥ ﻓﺮﺍﻭﺍﻧـﯽ ﺍﻓـﺮﺍﺩ‬
‫ﻫﺘﺮﻭﺯﻳﮕﻮﺕ ﺭﺍ ﺩﺭ ﺟﻤﻌﻴﺖ ﺑﺎﻻ ﺑﺮﺩ‪.‬‬

‫ﺷﮑﻞ ‪ -3‬ﻧﻤﻮﻧﻪ اي از ﻣﺤﺼﻮل ﻫﻀﻢ آﻧﺰﯾﻤﯽ ﺑﺮ روي ژل آﮐﺮﯾﻞ‬

‫آﻣﯿﺪ ‪%8‬‬

‫ﺗﺸﻜﺮ ﻭﻗﺪﺭﺩﺍﻧﻲ‬
‫ﺍﺯ ﺭﻳﺎﺳﺖ ﻣﺤﺘﺮﻡ ﻭ ﮐﺎﺭﮐﻨﺎﻥ ﻣﺮﮐﺰ ﺑﻴﻦ ﺍﻟﻤﻠﻠﻲ ﻋﻠـﻮﻡ ﻭ‬
‫ﺗﮑﻨﻮﻟــﻮﮊﻱ ﭘﻴــﺸﺮﻓﺘﻪ ﻭ ﻋﻠــﻮﻡ ﻣﺤﻴﻄــﻲ ﮐﺮﻣــﺎﻥ ﺟﻬــﺖ‬
‫ﺗﺨـــﺼﻴﺺ ﺍﻋﺘﺒـــﺎﺭ ﻭ ﺩﺭ ﺍﺧﺘﻴـــﺎﺭ ﻗـــﺮﺍﺭ ﺩﺍﺩﻥ ﺍﻣﮑﺎﻧـــﺎﺕ‬
‫ﺁﺯﻣﺎﻳﺸﮕﺎﻫﻲ ﺗﺸﮑﺮ ﻭ ﻗﺪﺭﺩﺍﻧﻲ ﻣﻲﺷﻮﺩ‪.‬‬ ‫ﺷﮑﻞ ‪ -2‬ﻧﻤﻮﻧﻪ اي از ﻣﺤﺼﻮﻻت ‪ PCR‬ﺑﺮ روي ژل آﮔﺎرز‬

‫ﺟﺪول ‪ -1‬ﻓﺮاواﻧﯽ ﻫﺎي ژﻧﯽ و ژﻧﻮﺗﯿﭙﯽ و ﺗﻌﺎدل ﻫﺎردي‪-‬وﯾﻨﺒﺮگ‬

‫ﻓﺮاواﻧﯽ ژﻧﻮﺗﯿﭙﯽ‬ ‫ﺗﻌﺪاد ﻣﻮرد اﻧﺘﻈﺎر‬ ‫ﺗﻌﺪاد ﻣﺸﺎﻫﺪه ﺷﺪه‬ ‫ژﻧﻮﺗﯿﭗ‬
‫‪0/02‬‬ ‫‪0/0936‬‬ ‫‪3‬‬ ‫‪MM‬‬
‫‪0/01‬‬ ‫‪7/8128‬‬ ‫‪2‬‬ ‫‪Mm‬‬
‫‪0/97‬‬ ‫‪142/0936‬‬ ‫‪145‬‬ ‫‪mm‬‬
‫‪0/0267‬‬ ‫ﻓﺮاواﻧﯽ ژن ‪M‬‬
‫‪0/9733‬‬ ‫ﻓﺮاواﻧﯽ ژن ‪m‬‬
‫‪94/5849‬‬ ‫آزﻣﻮن ﻣﺮﺑﻊ ﮐﺎ )‪ (χ2‬ﺑﺎ درﺟﻪ آزادي ‪1‬‬

‫ﻣﻨﺎﺑﻊ ﻣﻮﺭﺩ ﺍﺳﺘﻔﺎﺩﻩ‬

‫ﺳﻌﺎﺩﺕ ﻧﻮﺭﻱ ﻡ‪ .‬ﻭ ﺳﻴﺎﻩ ﻣﻨﺼﻮﺭ ﺹ‪ .۱۳۷۵ ،‬ﺍﺻﻮﻝ ﻧﮕﻬﺪﺍﺭﻱ ﻭﭘﺮﻭﺭﺵ ﮔﻮﺳﻔﻨﺪ‪ .‬ﺍﻧﺘﺸﺎﺭﺍﺕ ﺍﺷﺮﻓﻲ ﺗﻬﺮﺍﻥ‪.‬‬

‫ﻣﺴﻌﻮﺩﻱ ﻉ‪ ،‬ﻋﻤﺮﺍﻧﻲ ﺡ‪ ،‬ﻋﺒﺎﺳﻲ ﻉ‪ ،‬ﻧﺠﺎﺗﻲ ﺟﻮﺍﺭﻣﻲ ﺍﻟﻒ‪ ،‬ﻓﺮﻫﻨﮓ ﺥ‪ ،‬ﺍﺳﻤﺎﻋﻴﻞ ﺧﺎﻧﻴﺎﻥ ﺱ ﻭ ﺿﻴﺎﻳﻲ ﻑ‪ .۱۳۸۴ ،‬ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ‬
‫ﺗﮑﻨﻴﮏ ‪ PCR-SSCP‬ﺟﻬﺖ ﺑﺮﺭﺳﻲ ﭼﻨﺪ ﺷﮑﻠﻲ ﻫﺎﯼ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﻭ ﺍﺭﺗﺒـﺎﻁ ﺁﻥ ﺑـﺎ ﺻـﻔﺎﺕ ﺗﻮﻟﻴـﺪﻱ ﺩﺭ ﮔﻮﺳـﻔﻨﺪ‬
‫ﺑﻠﻮﭼﻲ‪ .‬ﭼﻬﺎﺭﻣﻴﻦ ﻫﻤﺎﻳﺶ ﻣﻠﻲ ﺑﻴﻮﺗﮑﻨﻮﻟﻮﮊﻱ ﺟﻤﻬﻮﺭﻱ ﺍﺳﻼﻣﻲ ﺍﻳﺮﺍﻥ‪.‬‬
 ١٣٨٨ ‫ ﺳﺎﻝ‬/١ ‫ ﺷﻤﺎﺭﻩ‬١٩ ‫ ﺟﻠﺪ‬/‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‬ ... ‫ ﻣﺤﻤﺪﺁﺑﺎﺩﯼ ﻭ‬،‫ﺻﻮﻓﯽ‬ ٨٨

‫ ﺍﻧﺘﺸﺎﺭﺍﺕ ﺟﻬﺎﺩ ﺳﺎﺯﻧﺪﮔﻲ‬.‫ ﮔﺰﺍﺭﺵ ﮔﻮﺳﻔﻨﺪ ﺳﻨﺠﺎﺑﻲ ﺩﺭ ﺑﺨﺶ ﺗﺤﻘﻴﻘﺎﺕ ﺩﺍﻣﭙﺮﻭﺭﻱ ﺍﺳﺘﺎﻥ ﮐﺮﻣﺎﻧﺸﺎﻩ‬.۱۳۷۸ ،‫ﻣﻮﻻﺋﻴﺎﻥ ﺡ‬
.۳۸ ‫ ﺹ‬.‫ﺍﺳﺘﺎﻥ ﮐﺮﻣﺎﻧﺸﺎﻩ‬

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Quantitative trait loci affecting growth and carcass composition of cattle segregating
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science papers and reports. 20:19-23.

Fahrankrug SC, Casas E, Keel JW and Smith TDL, 1999. Direct Genotyping of the Double-
muscling locus (mh) in piedmotese and Belgain Blue cattle by fluorescent PCR. Journal of
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Gerrard DE, Thrasher KH, Grant AL, Lemenager RP and Judge MD, 1991. Serum- induced
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(Ed. By JB Owen & RFE Axford), Oxford, Wallingford, UK. 467-478.

Jahnson PL, Mcewant JC, Dodds KG, Purchas RW and Blari HT, 2005. A directed search in the
region of GDF8 for quantitative trait loci affecting carcass trait in Texel sheep. Journal of
Animal Science. 83:1988-2000.

Jahnson PL, Mcewant JC, Dodds KG, Purchas RW and Blari HT, 2005. Meat quality traits were
unaffected by a quantitative trait locus affecting leg composition traits in Texel sheep.
Journal of Animal Science 83: 2729-2735.

Kobolak J and Gocza E, 2002. The role of the myostatin protein in meat quality. A review.
Archives of Animal Breeding 45:159-170.

Miller SA, Dykes DD and Polesky HF, 1998. A simple salting out procedure for extraction
DNA from human nucleated cells. Nucleic Acids Research 16:1255.

Smet SD, Webb EC, Claeys E, Uytterhaegen L and Demeyer DI, 2000. Effect of dietary energy
and protein levels on fatty acid composition of intramuscular fat in double- muscled Belgian
Blue bulls. Meat Science 56: 73-79.
 ٨٩ ... ‫ﺍﺭﺯﻳﺎﺑﯽ ﭼﻨﺪﺷﻜﻠﻲ ﮊﻥ ﻣﻴﻮﺳﺘﺎﺗﻴﻦ ﺩﺭ ﮔﻮﺳﻔﻨﺪ ﻧﮋﺍﺩ ﺳﻨﺠﺎﺑﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ‬

Timoty PL, Lopez-Corrales NL, Kappes SM and Sonstegard TS, 1997. Myostatin maps to the
interval containing the bovine mh locus. Mammalian Genome 8:742-744.

Wheeler TL, Shackelford SD, Casas E, Cundiff LV and Koohmavaie M, 2001. The effects of
piedomontese inheritance and myostatine genotype on the palatability of longissimus
thoracis, gluteus medius semimem branosus, and biceps femoris. Journal of animal science
79:3096- 3074.
 ‫ﻣﺠﻠﻪ ﭘﮋﻭﻫﺶﻫﺎﻱ ﻋﻠﻮﻡ ﺩﺍﻣﯽ‪ /‬ﺟﻠـﺪ ‪ ١٩‬ﺷـﻤﺎﺭﻩ‬ ‫ﺻﻮﻓﯽ‪ ،‬ﻣﺤﻤﺪﺁﺑﺎﺩﯼ ﻭ ‪...‬‬ ‫‪٩٠‬‬
‫‪ /١‬ﺳﺎﻝ ‪١٣٨٨‬‬

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