بررسی نقش جهش a501r در افزایش قدرت پردازش آنزیم dna پلیمراز pfu

‫ﺁﻧﺰیم ‬‫‪ⅾna‬ ﭘﻠﯿﻤﺮﺍﺯ‬ ‫‪pfu‬ﭘﺎیین‬ ‫ﺗﺮین ‬‫ﻧﺮﺥ‬ ‫ﺧﻄﺎ‬ ‫ﺩﺭ‬ ‫ﻓﺮﺁیند‬ ‫ﻫﻤﺎﻧﻨﺪ ﺳﺎﺯﯼ‬ ‫ﺩﺭ‬ ‫‪pⅽr‬ﺭﺍ‬ ‫ﺩﺍﺭﺩ‪.‬‬ ‫ﺍﻣﺎ‬‫ﻧﻘﻄﻪ‬ ‫ﺿﻌﻒ‬ ‫ﺍیﻦ‬ ‫ﺁﻧﺰیم‬ ‫ﭘﺎییین ‫ﺑﻮﺩﻥ‬ ‫ﻗﺪﺭﺕ‬ ‫ﭘﺮﺩﺍﺯﺵ‬ ‫ﺁﻥ‬ ‫ﺍﺳﺖ‬ ‫ﮐﻪ‬ ‫ﭘﺲ‬ ‫ﺍﺯ‬ ‫ﺍﻓﺰﻭﺩﻥ‬ ‫ﺗﻘﺮیبا ‬‫‪20‬‬ ‫ﻧﻮﮐﻠﺌﻮﺗﯿﺪ ‬‫ﺩﺭ‬ ‫ﺍﻧﺘﻬﺎﯼ‬ ‫ﭘﺮﺍیمر‬ ‫ﺍﺯ‬ ‫ﺭﻭﯼ‬ ‫ﺭﺷﺘﻪ‬ ‫‪ⅾna‬ ‫ﺍﻟ‬گو ﺑﻠﻨﺪ‬ ‫می‬ ‫ﺷﻮﺩ‪.‬‬ ‫ﺩﺭ‬ ‫ﺍین‬ ‫ﭘﮋﻭﻫﺶ‬ ‫ﻫﺪﻑ‬ ‫ﺍﻓﺰﺍیش ‫ﺗﻮﺍﻥ‬‫ﭘﺮﺩﺍﺯﺵ‬ ‫ﺁﻧﺰیم‬ ‫ﺑﺎ‬ ‫ﻃﺮﺍحی‬ ‫ﻣﻨﻄقی‬ ‫ﻭ‬ ‫ﺍیجاﺩ‬ ‫ﺟﻬﺶ‬ ‫ﻧﻘﻄە‬ ‫ﺍﯼ‬‫ ﺩﺭ‬ ‫ﺁﻧﺰیم‬ ‫ﺑﺎ‬ ‫ﮐﻤﺘﺮین‬ ‫ﻫﺰینه‬ ‫ﺁﻧﺘﺮﻭﭘﯽ‬ ‫ﻭ‬ ‫ﺁﻧﺘﺎﻟﭙﯽ ‬‫ﺍﺳﺖ‪ .‬ﺑرای‬ ‫ﻣﺮﺗﻔﻊ‬ ‫ﻧﻤﻮﺩﻥ‬ ‫ﻫﺪﻑ‬ ‫ﭘﮋﻭﻫﺶ‪،‬‬ ‫ﺍﺑﺘﺪﺍ‬ ‫ﻣﻘﺎیسه ‫ﺗﻮﺍلی ‫ﻭ‬ ‫ﺳﺎﺧﺘﺎﺭﯼ‬ ‫ﻣﯿﺎﻥ‬ ‫ﺁﻧﺰیم ‫ﻫﺎﯼ‬ ‫ﻫﻢ‬‫ ﺧﺎﻧﻮﺍﺩﻩ ‬‫ﺑﺎ‬ ‫ﺗﻮﺍﻥ‬ ‫ﭘﺮﺩﺍﺯﺵ‬ ‫ﺑﺎﻻ‬ ‫ﺍﻧﺠﺎﻡ‬ ‫ﺷﺪ‪،‬‬ ‫ﺳﭙﺲ‬ ‫ﺟﻬﺶ‬ ‫ﻣﻨﺎﺳﺐ‬ ‫ﺍﻧﺘﺨﺎﺏ‬ ‫ﮔﺮﺩید ‫ﻭ‬ ‫ﺷﺒیه ‫ﺳﺎﺯﯼ‬‫ ﺑﺮﺍﯼ‬ ‫ﻧﻤﻮنه ‬‫ﻃﺒیعی‬ ‫ﻭ‬ ‫ﺟﻬﺶ‬ ‫یاﻓﺘﻪ‬‫ﺍﻧﺠﺎﻡ‬ ‫ﺷﺪ‬ ‫ﻭ‬ ‫ﺧﺮﻭجی ‫ﺁﻥ‬ ‫ﻣﻮﺭﺩ‬ ‫ﺑﺮﺭسی ‬ ‫ﻗﺮﺍﺭ‬ ‫ﮔﺮﻓﺖ‬ ‫ﻭ‬ ‫ﻣﯿﺰﺍﻥ‬ ‫ﺍﻧﺮﮊﯼ‬ ‫ﺁﺯﺍﺩ‬ ‫ﺍﺗﺼﺎﻝ‬‫ﺁﻧﺰیم ‫ﻭ‬ ‫‪ⅾna‬ ﻧﺸﺎﻥ‬ ‫ﺩﺍﺩ‬ ‫ﮐﻪ‬ ‫ﻧﻤﻮﻧﻪ‬ ‫ﺟﻬﺶ‬ ‫یاﻓﺘﻪ‬‫ ﺍﺗﺼﺎﻝ‬ ‫ﺑﻬﺘﺮﯼ‬ ‫ﻧﺴﺒﺖ‬ ‫ﺑﻪ‬ ‫ﺁﻧﺰیم ‫ﻃﺒیعی  ‫ﺑﺎ‬ ‫‪ⅾna‬‬ ‫ﺑﺮﻗﺮﺍﺭ‬‫می ‫ﮐﻨﺪ‪.‬‬

the role of a501 mutation in processivity improvement of pfu dna polymerase

the pfu dna polymerase enzyme has the lowest error rate in the replication process during pcr. however, the drawback of this enzyme is its low processivity, which leads to halting after adding approximately 20 nucleotides at the end of the primer from the dna template strand. here we have studied and compared structurally and sequentially the pfu dna polymerase and dna polymerases in a same family with highest processivity and designed some mutations by rational design to improve processivity of the enzyme with the lowest entropy and enthalpy costs. in this investigation, conformational and behavioral changes have been studied by molecular dynamics simulation and for more precise information, free binding energy is calculated and the result shows that the mutant is willing to bind more tightly than native enzyme.